beta-Agarase I beta-琼脂糖酶 品牌:Nippon Gene


beta-Agarase I

beta-琼脂糖酶

品牌:Nippon Gene
CAS No.:37288-57-6
储存条件:-20℃
纯度:
产品编号

(生产商编号)

等级 规格 运输包装 零售价(RMB) 库存情况 参考值

316-02551

100 units

beta-Agarase I                                                      beta-琼脂糖酶            品牌:Nippon Gene


* 干冰运输、大包装及大批量的产品需酌情添加运输费用


* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动,恕不另行通知,确切详情请联系上海金畔生物科技有限公司。

Sambucus Sieboldiana Lectin-Agarose Suspension 无梗接骨木凝集素-琼脂糖 品牌:FUJIFILM Wako


Sambucus Sieboldiana Lectin-Agarose Suspension

无梗接骨木凝集素-琼脂糖

品牌:FUJIFILM Wako
CAS No.:N/A
储存条件:2-10℃
纯度:
产品编号

(生产商编号)

等级 规格 运输包装 零售价(RMB) 库存情况 参考值

191-10391

for Biochemistry 2 ml

Sambucus Sieboldiana Lectin-Agarose Suspension                                                      无梗接骨木凝集素-琼脂糖            品牌:FUJIFILM Wako


* 干冰运输、大包装及大批量的产品需酌情添加运输费用


* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动,恕不另行通知,确切详情请联系上海金畔生物科技有限公司。

Ulex Europaeus Lectin-1-Agarose Suspension 荆豆凝集素1-琼脂糖 品牌:FUJIFILM Wako


Ulex Europaeus Lectin-1-Agarose Suspension

荆豆凝集素1-琼脂糖

品牌:FUJIFILM Wako
CAS No.:
储存条件:2-10℃
纯度:
产品编号

(生产商编号)

等级 规格 运输包装 零售价(RMB) 库存情况 参考值

213-00891

for Biochemistry 2 ml

Ulex Europaeus Lectin-1-Agarose Suspension                                                      荆豆凝集素1-琼脂糖            品牌:FUJIFILM Wako


* 干冰运输、大包装及大批量的产品需酌情添加运输费用


* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动,恕不另行通知,确切详情请联系上海金畔生物科技有限公司。

Concanavalin A-Agarose 刀豆蛋白A-琼脂糖 品牌:FUJIFILM Wako


Concanavalin A-Agarose

刀豆蛋白A-琼脂糖

品牌:FUJIFILM Wako
CAS No.:
储存条件:2-10℃
纯度:
产品编号

(生产商编号)

等级 规格 运输包装 零售价(RMB) 库存情况 参考值

039-14101

for Biochemistry 10 ml

Concanavalin A-Agarose                                                      刀豆蛋白A-琼脂糖            品牌:FUJIFILM Wako


* 干冰运输、大包装及大批量的产品需酌情添加运输费用


* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动,恕不另行通知,确切详情请联系上海金畔生物科技有限公司。

Wheat Germ Lectin-Agarose 麦芽凝集素-琼脂糖 品牌:FUJIFILM Wako


Wheat Germ Lectin-Agarose

麦芽凝集素-琼脂糖

品牌:FUJIFILM Wako
CAS No.:
储存条件:2-10℃
纯度:
产品编号

(生产商编号)

等级 规格 运输包装 零售价(RMB) 库存情况 参考值

239-01191

for Biochemistry 2 ml

Wheat Germ Lectin-Agarose                                                      麦芽凝集素-琼脂糖            品牌:FUJIFILM Wako


* 干冰运输、大包装及大批量的产品需酌情添加运输费用


* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动,恕不另行通知,确切详情请联系上海金畔生物科技有限公司。

亲和性标签抗体结合琼脂糖珠 重组蛋白纯化用

亲和性标签抗体结合琼脂糖珠
重组蛋白纯化用

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

亲和性标签抗体结合琼脂糖珠                              重组蛋白纯化用

重组蛋白纯化用

亲和性标签抗体结合琼脂糖珠

亲和性标签抗体结合琼脂糖珠                              重组蛋白纯化用

亲和性标签抗体结合琼脂糖珠                              重组蛋白纯化用

◆亲和性标签抗体结合琼脂糖珠


亲和性标签抗体结合琼脂糖珠                              重组蛋白纯化用

可确认比A公司产品具有同等甚至以上的抗原回收性能!!

<使用载体量>
抗DYKDDDDK标签抗体琼脂糖珠(Wako): 20 μL / assay
A社 Affinity Beads: 20 μL /assay
<抗原添加量>
含DYKDDDDK标签融合蛋白的大肠杆菌细胞溶解液20 mg/assay
<免疫沉淀条件>
4℃、3 小时
<洗脱方法>
150 μg/mL DYKDDDDK肽 (产品编号:
044-30951) 20 μL /assay
→ 4℃、孵化30 分钟
添加20 μL / assay 的2%SDS sample buffer → 煮沸5分钟。

<SDS-PAGE>
样品电泳量:10 μL
<检测方法>
银染法
配制可过量表达DYKDDDDK标签融合蛋白(约19 kDa)的大肠杆菌裂解液,分别用本产品和A公司产品免疫沉淀后,用DYKDDDDK肽进行抗原洗脱。将得到的抗原样品用SDS-PAGE分离、银染色后,比较抗原回收率。

产品名称

子类
  克隆号

规格

产品编号

包装

DYK标签:DYKDDDK

抗DYKDDDDK标签抗体琼脂糖珠

Anti DYKDDDDK tag 

Antibody Beads

小鼠IgG2b

1E6

免疫化学用

012-22781

2 mL(Net 1 mL)

018-22783

10 mL(Net 5 mL)

016-22784

50 mL(Net 25 mL)

 识别N端、C端、Internal的DYKDDDDK标签
 使用载体:4%琼脂糖
 溶液组成:1×PBS (pH 7.4), 50 w/v% glycerol, 0.02 w/v%   sodium azide
 抗体结合量:7.5 mg/mL
 结合容量:1mL本产品可结合约1.0 mg的DYKDDDDK标签融合蛋白。

His标签:HHHHHH(6×Histidine)

抗6×组氨酸抗体琼脂糖珠

Anti 6×Histidine Antibody Beads

小鼠IgG3・κ
  28-75

免疫化学用

019-23391

2 mL(Net 1 mL)

015-23393

10 mL(Net 5 mL)

 可识别N端、C端的6×His tag
 使用载体:4%琼脂糖
 溶液组成:1×TBS (pH 7.4), 0.05 w/v% sodium azide
 抗体结合量:2.0 mg/mL
 结合容量:1 mL本产品可结合约1.0~2.0 mg的6×His标签融合蛋白

HA tag:YPYDVPDYA

抗HA抗体琼脂糖珠

Anti HA Antibody Beads

小鼠IgG1  

4B2

免疫化学用

014-23081

2 mL(Net 1 mL)

010-23083

10 mL(Net 5 mL)

 使用载体:4% 琼脂糖
 溶液组成:1×PBS (pH 7.4), 50% glycerol, 0.02 w/v% sodium azide
 抗体结合量:8.5 mg/mL
 结合容量:1 mL本品可结合约1.5 mg的HA标签融合蛋白

c-My标签:EQKLISEEDL

抗c-Myc抗体琼脂糖

Anti c-Myc Antibody Beads (10D11)

小鼠IgG1

10D11

免疫化学用

010-26501

200 μL

(Net 100 μL)

016-26503

2 mL(Net 1 mL)

014-26504

10 mL(Net 5 mL)

 使用担体:4% 琼脂糖
 溶液组成:1×PBS (pH 7.4), 50% glycerol, 0.02w/v% sodium azide
 抗体结合量:7.5mg/mL
 结合容量:1mL本产品可结合约0.9 mg的c-Myc tag融合蛋白

V5 tag:GKPIPNPLLGLDST

抗V5 tag抗体琼脂糖

Anti V5 tag Antibody Beads

小鼠IgG1
  10B5

免疫化学用

016-24381

2 mL(Net 1mL)

012-24383

10 mL(Net 5mL)

 使用载体:4%琼脂糖
 溶液组成:1×PBS (pH 7.4), 50% glycerol, 0.02w/v% sodium azide
 抗体结合量:7 mg/mL
 结合容量:1 mL本产品可结合约0.5 mg的V5标签融合蛋白

 

◆相关产品


内参照

下表为Western Blot实验中参照用单抗一览表。


产品名称

规格

产品编号

包装

抗β肌动蛋白单抗
  Anti β-Actin, Monoclonal Antibody

免疫化学用

013-24553

200 μg

011-24554

1 mg

017-24556

5 mg

抗β肌动蛋白单抗,过氧化物酶标记

Anti β-Actin, Monoclonal Antibody,   Peroxidase Conjugated

免疫化学用

017-24573

200 μL

抗α-微管蛋白, 单抗

Anti α-Tubulin, Monoclonal Antibody

免疫化学用

011-25034

10 μg

017-25031

200 μg

013-25033

1 mg

Anti β-Tubulin, Monoclonal Antibody
  抗β-微管蛋白, 单抗

免疫化学用

018-25044

10 μg

014-25041

200 μg

010-25043

1 mg

Anti GAPDH, Monoclonal Antibody
  抗GAPDH, 单抗

免疫化学用

016-25523

200 μg

014-25524

1 mg

010-25526

5 mg

Anti GAPDH, Monoclonal Antibody, Peroxidase   Conjugated
  抗GAPDH, 单抗, 过氧化酶结合

免疫化学用

015-25473

200 μL

 


蛋白酶抑制剂系列


产品名称

规格

产品编号

包装

蛋白酶抑制剂混合物试剂盒 I,无动物源成分(通用型)

Protease Inhibitor Cocktail Set I, Animal-derived-free (for general use) (×100), Lyophilized

通用型

165-26021

1 mL用

161-26023

1 mL用×5

蛋白酶抑制剂混合物试剂盒III,   DMSO溶液(不含EDTA)(×100)
  Protease Inhibitor Cocktail Set III, DMSO Solution   (EDTA free) (×100)

动物细胞

163-26061

1 mL

169-26063

1 mL×5

蛋白酶抑制剂混合物试剂盒V (无EDTA)(×100)
  Protease Inhibitor Cocktail Set V (EDTA free) (×100), Lyophilized

2D电泳

162-26031

1 mL

168-26033

1 mL×5

蛋白酶抑制剂混合物试剂盒 VI,DMSO溶液,用于植物(×100)
  Protease Inhibitor Cocktail Set VI, DMSO Solution (for Plant) (×100)

植物细胞

164-26091

1 mL

160-26093

1 mL×5

蛋白酶抑制剂混合物试剂盒VII,DMSO溶液,组氨酸标签蛋白(×100)
  Protease Inhibitor Cocktail Set VII, DMSO Solution   (for Histidine-tagged Protein) (×100)

His Tag 纯化

167-26101

1 mL

163-26103

1 mL×5

亲和标签比较表

亲和标签

TARGET

PA

6xHis

氨基酸序列

(YPGQ)5V

GVAMPGAEDDVV

HHHHHH

残基数

21

12

6

分子量 (kDa)

2.34

1.16

0.84

等电点(pI)

5.52

3.49

7.21

配体

TARGET tag

PA tag

Ni, Co, Zn, Cu

亲和珠回收

40% Propyleneglycol,
  1×TBS(pH7.5)

3M MgCl2, MES(pH6.0)

0.5M Imididazole(pH7.4)

单抗边界强度

(KD(M))

1.0×10-8

4.9×10-10

Ni-NTA:
  1.0×10-5~-6

单抗

Clone No.

P20.1(Mouse)

NZ-1(Rat)

Merck:HIS. H8(Mouse)

亲和力

++++

+++++

+++

提纯成本

+

+

+

多肽洗脱

+++

+++

Imidazole

背景

(培养上清)

+

+

+++++

背景

(细胞裂解物)

+

+

++++

亲和标签

DYKDDDDK

c-Myc

HA

GST

氨基酸序列

DYKDDDDK

EQKLISEEDL

YPYDVPDYA

残基数

8

10

9

218

分子量 (kDa)

1.01

1.20

1.10

26

等电点(pI)

3.97

4.00

3.56

配体

DYKDDDDK tag

c-Myc tag

HA tag

GST

亲和珠回收

Tris-HCl,
  Glycine Buffer(pH2~3.5)

Tris-HCl,
  Glycine Buffer(pH2~3.5)

Tris-HCl,
  Glycine Buffer(pH2~3.5)

Glutathione,
  Tris-HCl,
  Glycine Buffer(pH2~3.5)

单抗边界强度

(KD(M))

2.8×10-8

2.2×10-9

2.8×10-9

Glutathione:
  1.0×10-5~-6

单抗

Clone No.

M2(Mouse)

9E10(Mouse)

3F10(Mouse)
  HA-7(Rat)

Wako:5A7(Mouse)

亲和力

++++

++++

++++

++++

提纯成本

++

++

++

+

多肽洗脱

+++

++

++

Glutathione

背景

(培养上清)

++++

+++

+++

++++

背景

(细胞裂解物)

++++

++++

+++

++++

  本比较表为Wako独家调查的数据。样品、检测方法和手法的不同,特异性也会有所变化,因此无法保证各个亲和标签的功能。

 

※ 本页面产品仅供研究用,研究以外不可使用。


Ni-NTA琼脂糖 用于蛋白提取

Ni-NTA琼脂糖
用于蛋白提取

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

用于蛋白提取Ni-NTA琼脂糖                              用于蛋白提取

Ni-NTA琼脂糖

 


  本产品可用于亲和提取在N末端或C末端融合了带有六个组氨酸残基的6×His标签的重组蛋白。它用于提取多种蛋白质,如可溶性蛋白质,不溶性蛋白质,膜蛋白质等等。溶出时无需使用多肽,以低成本和简单的操作即可提取目的蛋白。

 


◆特点


● 具有高性价比

● 可再生利用



◆产品规格

 凝胶基质

 6% 交联琼脂糖

 配体

 次氮基三乙酸(NTA)

 Beads size

 50-150 μm

 蛋白结合容量

 >50 mg/mL 凝胶

*蛋白结合容量会因目的蛋白不同而有所差异。

 


数据


Ni-NTA琼脂糖                              用于蛋白提取


可以与其他公司的产品回收等量的蛋白质。


<实验条件>

① 通过超声破碎法从大肠杆菌中回收6×组氨酸融合GFP,作为精制前样本。

② 将Ni–NTA柱连接到液相色谱系统。

③ 用150 mL Running Buffer*1平衡色谱柱。

④ 上样精制前样本。

⑤ 用100 mL Running Buffer清洗。

⑥ 用150 mL Elution Buffer*2洗脱。

⑦ 用100 mL Running Buffer清洗。

⑧ 进行SDS-PAGE、CBB染色。


*1 Running Buffer:20 mmol/L Sodium Phosphate(pH 7.4),

          500 mmol/L Sodium Chloride, 

          10 mmol/L Imidazole

*2 Elution Buffer:20 mmol/L Sodium Phosphate(pH 7.4),

           500 mmol/L Sodium Chloride,

                                10~500 mmol/L Imidazole


具体实验条件请根据精制的目的蛋白进行检讨。



◆产品列表

产品编号

产品名称

规格

包装

147-09761

Ni-NTA Agarose
Ni-NTA琼脂糖

基因研究用

2 mL(Net 1 mL)

143-09763

10 mL(Net 5 mL)

141-09764

100 mL(Net 50 mL)



相关产品


预装型


Ni-NTA琼脂糖                              用于蛋白提取


本产品是预装柱产品。可直接用于液相色谱系统。

产品编号

产品名称

规格

包装

146-09731

Ni-NTA Cartridge
Ni-NTA预装柱

基因研究用

1EA(5 mL)

142-09733

1EA(5 mL)×5


高性能型


具有高蛋白结合能力的高性能型Ni-NTA琼脂糖。

产品编号

产品名称

规格

包装

149-09684

Ni-NTA Agarose HP
Ni-NTA琼脂糖 高性能型

基因研究用

2 mL(Net 1 mL)

145-09681

10 mL(Net 5 mL)

141-09683

100 mL(Net 50 mL)

※ 本页面产品仅供研究用,研究以外不可使用。

试剂怎么用——琼脂糖


试剂怎么用——琼脂糖

试剂怎么用——琼脂糖

试剂怎么用——琼脂糖

即使是为同一目的所用的试剂种类繁多,同一物质也会有不同的浓度、纯度、规格。

因此,经常有研究人员向我们问到“不知道哪一种适合自己的实验”等。

为了解决这样的问题,小光老师将介绍一些正确区分使用FUJIFILM Wako品牌试剂的方法。

◆琼脂糖

应用于核酸电泳的琼脂糖是从石花菜、江蓠等红藻中所提取的高纯度琼脂。

琼脂糖虽在分子生物学实验中较为常见,但其实种类繁多。为选择适合实验目的的琼脂糖,需检查以下项目。


● 凝胶强度

琼脂糖凝胶破裂所需的载荷用(g/cm2)来表示。标准琼脂糖约为≧1,200 g/cm2(浓度1.5%,Nippon Gene Agarose S)。分离高分子核酸时,需要使用低浓度琼脂糖,因此使用高凝胶强度的琼脂糖可更有效地制备凝胶。反之,在分离低分子核酸时,则使用凝胶强度低的琼脂糖。


● 熔点

琼脂糖凝胶融化的温度。标准琼脂糖的熔点在90°C左右,可通过引入羟乙基来降低熔点。这种低熔点型琼脂糖比双链DNA的变性温度要低,加热至65°C左右即会熔化,因此应用于DNA回收等。低熔点琼脂糖的胶凝化温度也比其他琼脂糖低。


● 硫酸含量

应用于电泳的琼脂糖由琼脂糖和琼脂胶组成,杂质琼脂胶中含有硫酸基团。因此,琼脂糖标准品中的硫酸含量表示琼脂糖的纯化程度,高纯度琼脂糖的硫酸含量表示为≤0.1-0.2%。


● 电渗度

琼脂糖因带有硫酸基等而带负电,接触的缓冲液带正电,两者极性相反。在这种状态下进行电泳时,带正电的缓冲溶液向阴极移动。这种现象称为电渗,也是琼脂糖凝胶电泳中迁移缓慢的原因。因此,一般使用电渗度低的琼脂糖进行电泳。

Nippon Gene Agarose 产品系列及正确使用方法

产品名

产品编号

凝胶强度

熔点

胶凝温度

硫酸含量

电渗度

分离范围

使用浓度范围

用途

Agarose S

318-01195

≧1,200g/cm2

(1.5%)

≦90℃(1.5%)

37-39℃(1.5%)

≦0.1%

≦0.1

0.5-30kbp

0.5-2%

标准琼脂糖。可制备的凝胶浓度范围广,成本低。

Agarose HS

312-01431

≧1,600g/cm2

(1.5%)

≦93℃(1.5%)

37-39℃(1.5%)

≦0.1%

≦0.1

0.5-30kbp

0.5-2%

凝胶强度高,Southern印迹的理想选择。

Agarose H

312-01431

≧2,600g/cm2

(1.5%)

Boil(1.5%)

37-39℃(1.5%)

≦0.2%

≦0.2

1-200kbp

0.2-1%

可制备高凝胶强度、低浓度的凝胶。分离高分子量核酸的理想选择。

Agarose XP

316-06515

≧450g/cm2

(1.5%)

≦65℃(1.5%)

≦30℃(1.5%)

≦0.1%

≦0.1

0.01-20kbp

1-4%

低熔点琼脂糖。凝胶温度低,容易回收核酸。

Agarose 21

319-03244

≧800g/cm2

(3%)

≦85℃(3%)

34-38℃(3%)

≦0.1%

≦0.1

0.01-1.0kbp

2-5%

溶解性优异,可制备高浓度凝胶。分离小分子核酸的理想选择。

Agarose X

313-02681

≧1,200g/cm2

(4%)

≦92℃(4%)

31-39℃(4%)

≦0.15%

0.06-0.14

0.01-1.0kbp

2-6%

溶解性优异,可制备高浓度凝胶。凝胶强度高于Agarose   21。

【参考文献】


1)電気泳動学会 編:「新版 電気泳動実験法」(文光堂)(1989).

2)大藤道衛 編:「電気泳動なるほどQ&A」(羊土社)(2007).

3)Green, R. M. and Sambrook, J.: ”Molecular Cloning A Laboratory Manual, 4th ed.”, Cold Spring Harbor Laboratory Press(2012).

Phos-tag™ 琼脂糖枪头 Phos-tag™ Tip

Phos-tag™ 琼脂糖枪头
Phos-tag™ Tip

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

Phos-tag™ 琼脂糖枪头Phos-tag™ 琼脂糖枪头                              Phos-tag™ Tip

Phos-tag™ Tip

  可特异性捕捉磷酸基团的功能性分子“Phos-tag ™”的磷酸化肽纯化用枪头。

  枪头里含有 Phos-tag ™ 琼脂糖,是即开即用的前处理工具,适用于生理状态下低分子量磷酸化分子(核酸和多肽等)的分离和富集。


原理


把 Phos-tag™ Tip 装在注射器(购买产品附送)上使用。


                                   

Phos-tag™ 琼脂糖枪头                              Phos-tag™ Tip

 

                                                           

Phos-tag ™ Tip 的结构


Phos-tag™ 琼脂糖枪头                              Phos-tag™ Tip

优点、特色


 操作时间少于30分钟 

 高回收率可重复利用

 无需昂贵仪器

 缓冲液为生理 pH 条件下


案例、应用


分离磷酸化肽使用例


Phos-tag™ 琼脂糖枪头                              Phos-tag™ Tip



Phos-tag™ 系列

磷酸化蛋白新方法!

  Phos-tag™是一种能与磷酸离子特异性结合的功能性分子。它可用于磷酸化蛋白的分离(Phos-tag™ Acrylamide)、Western Blot 检测(Phos-tag™ Biotin)、蛋白纯化 (Phos-tag™Agarose)及质谱分析 MALDI-TOF/MS (Phos-tag™ Mass Analytical Kit)。


◆Phos-tag™ 的基本结构

Phos-tag™ 琼脂糖枪头                              Phos-tag™ Tip



原理:


Phos-tag™ 琼脂糖枪头                              Phos-tag™ Tip



特点:


●   与 -2 价磷酸根离子的亲和性和选择性高于其它阴离子

●   在 pH 5-8 的生理环境下生成稳定的复合物

相关应用:


Phos-tag™ 琼脂糖枪头                              Phos-tag™ Tip

相关产品:

 产品名称

 用  途

 Phos-tag™ Acrylamide

 分离:SDS – PAGE 分离不同磷酸化水平的蛋白

 SuperSep Phos-tag™

 分离:预制胶中含有 50 μM Phos-tag™ Acrylamide

 Phos-tag™ Biotin

 检测:代替 Western Blot 检测中的磷酸化抗体

 Phos-tag™ Agarose

 纯化:通用柱层析,纯化磷酸化蛋白

 Phos-tag™ Mass

 Analytical Kit

 分析:用于质谱 MALDI-TOF/MS 分析,提高磷酸化分子的检测灵敏度


phos-tag™ 由日本广岛大学研究生院医齿药学综合研究科医药分子功能科学研究室开发。

更多产品信息,请点击:

Phos-tag™ 琼脂糖枪头                              Phos-tag™ Tip

Phos-tag 第6版说明书

Phos-tag™ 琼脂糖枪头                              Phos-tag™ Tip

Phos-tag系列 ver. 8


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·  Transcriptional Profile during Deoxycholate-Induced Sporulation in a Clostridium perfringens Isolate Causing Foodborne Illness[J]. Applied and environmental microbiology, 2016, 82(10): 2929-2942,Yasugi M, Okuzaki D, Kuwana R, et al.

·  Timely Closure of the Prospore Membrane Requires SPS1 and SPO77 in Saccharomyces cerevisiae[J]. Genetics, 2016: genetics. 115.183939,Paulissen S M, Slubowski C J, Roesner J M, et al.

·  DDK dependent regulation of TOP2A at centromeres revealed by a chemical genetics approach[J]. Nucleic Acids Research, 2016: gkw626,Wu K Z L, Wang G N, Fitzgerald J, et al.

·  OVATE Family Protein 8 Positively Mediates Brassinosteroid Signaling through Interacting with the GSK3-like Kinase in Rice[J]. PLoS Genet, 2016, 12(6): e1006118,Yang C, Shen W, He Y, et al.

·  Epithelial Sel1L is required for the maintenance of intestinal homeostasis[J]. Molecular biology of the cell, 2016, 27(3): 483-490, Sun S, Lourie R, Cohen S B, et al.

·  Effect of Sodium Dodecyl Sulfate Concentration on Supramolecular Gel Electrophoresis[J]. ChemNanoMat, 2016,Tazawa S, Kobayashi K, Yamanaka M.

·  Intergenic VNTR Polymorphism Upstream of rocA Alters Toxin Production and Enhances Virulence in Streptococcus pyogenes[J]. Infection and immunity, 2016: IAI. 00258-16,Zhu L, Olsen R J, Horstmann N, et al.

·  Ajuba Phosphorylation by CDK1 Promotes Cell Proliferation and Tumorigenesis[J]. Journal of Biological Chemistry, 2016: jbc. M116. 722751,Chen X, Stauffer S, Chen Y, et al.

·  Editorial: International Plant Proteomics Organization (INPPO) World Congress 2014[J]. Frontiers in Plant Science, 2016, 7,Heazlewood J L, Jorrín-Novo J V, Agrawal G K, et al.

·  Phosphoinositide kinase signaling controls ER-PM cross-talk[J]. Molecular biology of the cell, 2016, 27(7): 1170-1180,Omnus D J, Manford A G, Bader J M, et al.

·  A multiple covalent crosslinked soft hydrogel for bioseparation[J]. Chemical Communications, 2016, 52(15): 3247-3250,Liu Z, Fan L, Xiao H, et al.

·  Advances in crop proteomics: PTMs of proteins under abiotic stress[J]. Proteomics, 2016, 16(5): 847-865,Wu X, Gong F, Cao D, et al.

·  Cyclin-Dependent Kinase Co-Ordinates Carbohydrate Metabolism and Cell Cycle in S. cerevisiae[J]. Molecular cell, 2016, 62(4): 546-557,Zhao G, Chen Y, Carey L, et al.

·  Carbon Monoxide Gas Is Not Inert, but Global, in Its Consequences for Bacterial Gene Expression, Iron Acquisition, and Antibiotic Resistance[J]. Antioxidants & redox signaling, 2016,Wareham L K, Begg R, Jesse H E, et al.

·  Two-layer regulation of PAQR3 on ATG14-linked class III PtdIns3K activation upon glucose starvation[J]. Autophagy, 2016: 1-2,Xu D, Wang Z, Chen Y.

·  Regulation of sphingolipid biosynthesis by the morphogenesis checkpoint kinase Swe1[J]. Journal of Biological Chemistry, 2016, 291(5): 2524-2534,Chauhan N, Han G, Somashekarappa N, et al.

·  PAX5 tyrosine phosphorylation by SYK co-operatively functions with its serine phosphorylation to cancel the PAX5-dependent repression of BLIMP1: A mechanism for antigen-triggered plasma cell differentiation[J]. Biochemical and biophysical research communications, 2016, 475(2): 176-181,Inagaki Y, Hayakawa F, Hirano D, et al.

·  A Combined Computational and Genetic Approach Uncovers Network Interactions of the Cyanobacterial Circadian Clock[J]. Journal of Bacteriology, 2016: JB. 00235-16,Boyd J S, Cheng R R, Paddock M L, et al.

·  HuR mediates motility of human bone marrow-derived mesenchymal stem cells triggered by sphingosine 1-phosphate in liver fibrosis[J]. Journal of Molecular Medicine, 2016: 1-14,Chang N, Ge J, Xiu L, et al.

·  Combined replacement effects of human modified β-hexosaminidase B and GM2 activator protein on GM2 gangliosidoses fibroblasts[J]. Biochemistry and Biophysics Reports, 2016,Kitakaze K, Tasaki C, Tajima Y, et al.

·  Roseotoxin B Improves Allergic Contact Dermatitis through a Unique Anti-inflammatory Mechanism Involving Excessive Activation of Autophagy in Activated T-Lymphocytes[J]. Journal of Investigative Dermatology, 2016,Wang X, Hu C, Wu X, et al.


References on Phos-tag™ Chemistry

  • Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of phosphorylated compounds using a novel phosphate capture moleculeRapid Communications of Mass Spectrometry17, 2075-2081 (2003), H. Takeda, A. Kawasaki, M. Takahashi, A. Yamada, and T. Koike 

  • Recognition of phosphate monoester dianion by an alkoxide-bridged dinuclear zinc (II) complexDalton Transactions, 1189-1193 (2004), E. Kinoshita, M. Takahashi, H. Takeda, M. Shiro, and T. Koike

  • Quantitative analysis of lysophosphatidic acid by time-of-flight mass spectrometry using a phosphate capture molecule, Journal of Lipid Research45, 2145-2150 (2004), T. Tanaka, H. Tsutsui, K. Hirano, T. Koike, A. Tokumura, and K. Satouchi

  •  Production of 1,2-Didocosahexaenoyl Phosphatidylcholine by Bonito Muscle Lysophosphatidylcholine/TransacylaseJournal of Biochemistry,136, 477-483 (2004), K. Hirano, H. Matsui, T. Tanaka, F. Matsuura, K. Satouchi, and T. Koike

  • Novel immobilized zinc(II) affinity chromatography for phosphopeptides and phosphorylated proteins, Journal of Separation Science, 28, 155-162 (2005), E. Kinoshita, A. Yamada, H. Takeda, E. Kinoshita-Kikuta, and T. Koike

  • Detection and Quantification of On-Chip Phosphorylated Peptides by Surface Plasmon Resonance Imaging Techniques Using a Phosphate Capture MoleculeAnalytical Chemistry77, 3979-3985 (2005), K. Inamori, M. Kyo, Y. Nishiya, Y. Inoue, T. Sonoda, E. Kinoshita, T. Koike, and Y. Katayama

  • Phosphate-binding tag: A new tool to visualize phosphorylated proteins, Molecular & Cellular Proteomics, 5, 749-757 (2006), E. Kinoshita, E. Kinoshita-Kikuta, K. Takiyama, and T. Koike

  • Enrichment of phosphorylated proteins from cell lysate using phosphate-affinity chromatography at physiological pHProteomics, 6, 5088-5095 (2006), E. Kinoshita-Kikuta, E. Kinoshita, A. Yamada, M. Endo, and T. Koike

  • Separation of a phosphorylated histidine protein using phosphate affinity polyacrylamide gel electrophoresis, Analytical Biochemistry360, 160-162 (2007), S. Yamada, H. Nakamura, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, and Y. Shiro

  • Label-free kinase profiling using phosphate-affinity polyacrylamide gel electrophresisMolecular & Cellular Proteomics, 6, 356-366 (2007), E. Kinoshita-Kikuta, Y. Aoki, E. Kinoshita, and T. Koike

  • A SNP genotyping method using phosphate-affinity polyacrylamide gel electrophoresis, Analytical Biochemistry361, 294-298 (2007), E. Kinoshita, E. Kinoshita-Kikuta, and T. Koike (The phosphate group at DNA-terminal is efficiently captured by Zn2+.Phos-tag.)

  • Identification on Membrane and Characterization of Phosphoproteins Using an Alkoxide-Bridged Dinuclear Metal Complex as a Phosphate-Binding Tag MoleculeJournal of Biomolecular Techniques18, 278-286 (2007), T. Nakanishi, E. Ando, M. Furuta, E. Kinoshita, E. Kikuta-Kinoshita, T. Koike, S. Tsunasawa, and O. Nishimura

  • A mobility shift detection method for DNA methylation analysis using phosphate affinity polyacrylamide gel electrophoresisAnalytical Biochemistry378, 102-104 (2008), E. Kinoshita-Kikuta, E. Kinoshita, and T. Koike

  • Separation of phosphoprotein isotypes having the same number of phosphate groups using phosphate- affinity SDS-PAGEProteomics, 8, 2994-3003 (2008), E. Kinoshita, E. Kinoshita-Kikuta, M. Matsubara, S. Yamada, H. Nakamura, Y. Shiro, Y. Aoki, K. Okita, and T. Koike

  • FANCI phosphorylation functions as a molecular switch to turn on the Fanconi anemia pathwayNature Structural & Molecular Biology15, 1138-1146 (2008), M. Ishiai, H. Kitao, A. Smogorzewska, J. Tomida, A. Kinomura, E. Uchida, A. Saberi, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, S. Tashiro, S. J. Elledge, and M. Takata

  • to Page top

  • Two-dimensional phosphate affinity gel electrophoresis for the analysis of phosphoprotein isotypes Electrophoresis30, 550-559 (2009), E. Kinoshita, E. Kinoshita-Kikuta, M. Matsubara, Y. Aoki, S. Ohie, Y. Mouri, and T. Koike

  • Formation of lysophosphatidic acid, a wound-healing lipid, during digestion of cabbage leavesBioscience, Biotechnology, and Biochemistry,73, 1293-300 (2009), T. Tanaka, G. Horiuchi, M. Matsuoka, K. Hirano, A. Tokumura, T. Koike, and K. Satouchi

  • A Phos-tag-based fluorescence resonance energy transfer system for the analysis of the dephosphorylation of phosphopeptidesAnalytical Biochemistry388, 235-241, (2009), K. Takiyama, E. Kinoshita, E. Kinoshita-Kikuta, Y. Fujioka, Y. Kubo, and T. Koike

  • Phos-tag beads as an immunoblotting enhancer for selective detection of phosphoproteins in cell lysatesAnalytical Biochemistry389, 83-85, (2009), E. Kinoshita-Kikuta, E. Kinoshita, and T. Koike

  • Mobility shift detection of phosphorylation on large proteins using a Phos-tag SDS-PAGE gel strengthened with agaroseProteomics9, 4098- 4101 (2009), E. Kinoshita, E. Kinoshita-Kikuta, H. Ujihara, and T. Koike

  • Separation and detection of large phosphoproteins using Phos-tag SDS-PAGENature Protocols4, 1513-1521 (2009), E. Kinoshita, E. Kinoshita-Kikuta, and T. Koike

  • A clean-up technology for the simultaneous determination of lysophosphatidic acid and sphingosine-1-phosphate by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using a phosphate-capture molecule, Phos-tagRapid Communications in Mass Spectrometry24, 1075-1084 (2010), J. Morishige, M. Urikura, H. Takagi, K. Hirano, T. Koike, T. Tanaka, and K. Satouchi

  • Genotyping and mapping assay of single-nucleotide polymorphisms in CYP3A5 using DNA-binding zinc(II) complexesClinical Biochemistry43, 302-306 (2010), E. Kinoshita, E. Kinoshita-Kikuta, H. Nakashima, and T. Koike

  • The DNA-binding activity of mouse DNA methyltransferase 1 is ragulated phosphorylation with casein kinase 1σ/εBiochemical Journal427, 489-497 (2010), Y. Sugiyama, N. Hatano, N. Sueyoshi, I. Suetake, S. Tajima, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, and I. Kameshita


产品列表
产品编号 产品名称 产品规格 产品等级 备注
387-07321 Phos-tag™ Tip 
Phos-tag™ 琼脂糖枪头
8个

Phos-tag™ 琼脂糖 Phos-tag™ Agarose

Phos-tag™ 琼脂糖
Phos-tag™ Agarose

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

Phos-tag™ AgarosePhos-tag™ 琼脂糖                              Phos-tag™ Agarose

亲和层析纯化磷酸化蛋白

填入色谱柱中使用。可分离、纯化、浓缩磷酸化蛋白。不使用界面活性剂、还原剂,可得到状态近似生物体内的磷酸化蛋白。

Phos-tag™ 琼脂糖                              Phos-tag™ Agarose

原理

Phos-tag™ 琼脂糖                              Phos-tag™ Agarose

Phos-tag™ 琼脂糖                              Phos-tag™ Agarose

优点、特色

● 无需使用还原剂或表面活性剂即可纯化

● 与亲和层析方法类似。

● 可在1小时内纯化磷酸化蛋白。

● Phos-tag™ Agarose 捕获结合到 Tyr、Thr、Ser、Asp、His 等氨基酸、糖类、脂类上的无机磷酸根和大量二价磷酸根。

● 可在生理条件下(pH7.5)捕捉蛋白。

● 纯化后的产物可用于 Co-IP 实验和其他蛋白活性实验。

案例、应用:

【使用例子:A431 裂解液中的磷酸化蛋白的纯化】

把Phos-tag™ 填充到柱里,再加上 A431 裂解液。

SYPRO Ruby 染色(左图)再使用 Anti-p Tyr 抗体进行免疫印迹(右图),检测出结果。

结果确认磷酸化蛋白浓缩在柱吸附层里。

M:分子量标记

Lane 1:未吸附层

Lane 2:吸附层

Lane 3:柱清洗层

Phos-tag™ 琼脂糖                              Phos-tag™ Agarose

Phos-tag™ 系列

磷酸化蛋白新方法!

  Phos-tag™ 是一种能与磷酸离子特异性结合的功能性分子。它可用于磷酸化蛋白的分离(Phos-tag™ Acrylamide)、Western Blot 检测(Phos-tag™ Biotin)、蛋白纯化 (Phos-tag™ Agarose)及质谱分析 MALDI-TOF/MS (Phos-tag™ Mass Analytical Kit)。


Phos-tag™ 的基本结构


Phos-tag™ 琼脂糖                              Phos-tag™ Agarose


特点


与 -2 价磷酸根离子的亲和性和选择性高于其它阴离子

在 pH 5-8 的生理环境下生成稳定的复合物

相关应用


Phos-tag™ 琼脂糖                              Phos-tag™ Agarose

相关产品

 产品名称

 用  途

 Phos-tag™ Acrylamide

 分离SDS – PAGE 分离不同磷酸化水平的蛋白

 SuperSep Phos-tag™

 分离预制胶中含有 50 μM Phos-tag™ Acrylamide

 Phos-tag™ Biotin

 检测代替 Western Blot 检测中的磷酸化抗体

 Phos-tag™ Agarose

 纯化通用柱层析,纯化磷酸化蛋白

 Phos-tag™ Mass

 Analytical Kit

 分析:用于质谱 MALDI-TOF/MS 分析,提高磷酸化分子的检测灵敏度


phos-tag™ 由日本广岛大学研究生院医齿药学综合研究科医药分子功能科学研究室开发。

更多产品信息,请点击:http://phos-tag.jp

Phos-tag™ 琼脂糖                              Phos-tag™ Agarose

Phos-tag 第6版说明书

Phos-tag™ 琼脂糖                              Phos-tag™ Agarose

Phos-tag系列 ver. 8



Phos-tag™ 琼脂糖                              Phos-tag™ Agarose

说明书

【参考文献】


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·  Sequential domain assembly of ribosomal protein S3 drives 40S subunit maturation[J]. Nature communications, 2016, 7,Mitterer V, Murat G, Réty S, et al.

·  Phos-tag analysis of Rab10 phosphorylation by LRRK2: a powerful assay for assessing kinase function and inhibitors[J]. Biochemical Journal, 2016: BCJ20160557,Ito G, Katsemonova K, Tonelli F, et al.

·  Analysis of phosphorylation of the myosin targeting subunit of smooth muscle myosin light chain phosphatase by Phos-tag SDS-PAGE[J]. The FASEB Journal, 2016, 30(1 Supplement): 1209.1-1209.1,Walsh M P, MacDonald J A, Sutherland C.

·  Using Phos-Tag in Western Blotting Analysis to Evaluate Protein Phosphorylation[J]. Kidney Research: Experimental Protocols, 2016: 267-277,Horinouchi T, Terada K, Higashi T, et al.

·  The Abundance of Nonphosphorylated Tau in Mouse and Human Tauopathy Brains Revealed by the Use of Phos-Tag Method[J]. The American journal of pathology, 2016, 186(2): 398-409,Kimura T, Hatsuta H, Masuda-Suzukake M, et al.

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·  Validation of Cis and Trans Modes in Multistep Phosphotransfer Signaling of Bacterial Tripartite Sensor Kinases by Using Phos-Tag SDS-PAGE[J]. PloS one, 2016, 11(2): e0148294,Kinoshita-Kikuta E, Kinoshita E, Eguchi Y, et al.

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·  Luteinizing Hormone Causes Phosphorylation and Activation of the cGMP Phosphodiesterase PDE5 in Rat Ovarian Follicles, Contributing, Together with PDE1 Activity, to the Resumption of Meiosis[J]. Biology of reproduction, 2016: biolreprod. 115.135897,Egbert J R, Uliasz T F, Shuhaibar L C, et al.

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References on Phos-tag™ Chemistry

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of phosphorylated compounds using a novel phosphate capture moleculeRapid Communications of Mass Spectrometry17, 2075-2081 (2003), H. Takeda, A. Kawasaki, M. Takahashi, A. Yamada, and T. Koike 

Recognition of phosphate monoester dianion by an alkoxide-bridged dinuclear zinc (II) complexDalton Transactions, 1189-1193 (2004), E. Kinoshita, M. Takahashi, H. Takeda, M. Shiro, and T. Koike

Quantitative analysis of lysophosphatidic acid by time-of-flight mass spectrometry using a phosphate capture molecule, Journal of Lipid Research45, 2145-2150 (2004), T. Tanaka, H. Tsutsui, K. Hirano, T. Koike, A. Tokumura, and K. Satouchi

Production of 1,2-Didocosahexaenoyl Phosphatidylcholine by Bonito Muscle Lysophosphatidylcholine/TransacylaseJournal of Biochemistry,136, 477-483 (2004), K. Hirano, H. Matsui, T. Tanaka, F. Matsuura, K. Satouchi, and T. Koike

Novel immobilized zinc(II) affinity chromatography for phosphopeptides and phosphorylated proteins, Journal of Separation Science, 28, 155-162 (2005), E. Kinoshita, A. Yamada, H. Takeda, E. Kinoshita-Kikuta, and T. Koike

Detection and Quantification of On-Chip Phosphorylated Peptides by Surface Plasmon Resonance Imaging Techniques Using a Phosphate Capture MoleculeAnalytical Chemistry77, 3979-3985 (2005), K. Inamori, M. Kyo, Y. Nishiya, Y. Inoue, T. Sonoda, E. Kinoshita, T. Koike, and Y. Katayama

Phosphate-binding tag: A new tool to visualize phosphorylated proteins, Molecular & Cellular Proteomics, 5, 749-757 (2006), E. Kinoshita, E. Kinoshita-Kikuta, K. Takiyama, and T. Koike

Enrichment of phosphorylated proteins from cell lysate using phosphate-affinity chromatography at physiological pHProteomics, 6, 5088-5095 (2006), E. Kinoshita-Kikuta, E. Kinoshita, A. Yamada, M. Endo, and T. Koike

Separation of a phosphorylated histidine protein using phosphate affinity polyacrylamide gel electrophoresis, Analytical Biochemistry360, 160-162 (2007), S. Yamada, H. Nakamura, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, and Y. Shiro

Label-free kinase profiling using phosphate-affinity polyacrylamide gel electrophresisMolecular & Cellular Proteomics, 6, 356-366 (2007), E. Kinoshita-Kikuta, Y. Aoki, E. Kinoshita, and T. Koike

A SNP genotyping method using phosphate-affinity polyacrylamide gel electrophoresis, Analytical Biochemistry361, 294-298 (2007), E. Kinoshita, E. Kinoshita-Kikuta, and T. Koike (The phosphate group at DNA-terminal is efficiently captured by Zn2+.Phos-tag.)

Identification on Membrane and Characterization of Phosphoproteins Using an Alkoxide-Bridged Dinuclear Metal Complex as a Phosphate-Binding Tag MoleculeJournal of Biomolecular Techniques18, 278-286 (2007), T. Nakanishi, E. Ando, M. Furuta, E. Kinoshita, E. Kikuta-Kinoshita, T. Koike, S. Tsunasawa, and O. Nishimura

A mobility shift detection method for DNA methylation analysis using phosphate affinity polyacrylamide gel electrophoresisAnalytical Biochemistry378, 102-104 (2008), E. Kinoshita-Kikuta, E. Kinoshita, and T. Koike

Separation of phosphoprotein isotypes having the same number of phosphate groups using phosphate- affinity SDS-PAGEProteomics, 8, 2994-3003 (2008), E. Kinoshita, E. Kinoshita-Kikuta, M. Matsubara, S. Yamada, H. Nakamura, Y. Shiro, Y. Aoki, K. Okita, and T. Koike

FANCI phosphorylation functions as a molecular switch to turn on the Fanconi anemia pathwayNature Structural & Molecular Biology15, 1138-1146 (2008), M. Ishiai, H. Kitao, A. Smogorzewska, J. Tomida, A. Kinomura, E. Uchida, A. Saberi, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, S. Tashiro, S. J. Elledge, and M. Takata

Two-dimensional phosphate affinity gel electrophoresis for the analysis of phosphoprotein isotypes Electrophoresis30, 550-559 (2009), E. Kinoshita, E. Kinoshita-Kikuta, M. Matsubara, Y. Aoki, S. Ohie, Y. Mouri, and T. Koike

Formation of lysophosphatidic acid, a wound-healing lipid, during digestion of cabbage leavesBioscience, Biotechnology, and Biochemistry,73, 1293-300 (2009), T. Tanaka, G. Horiuchi, M. Matsuoka, K. Hirano, A. Tokumura, T. Koike, and K. Satouchi

A Phos-tag-based fluorescence resonance energy transfer system for the analysis of the dephosphorylation of phosphopeptidesAnalytical Biochemistry388, 235-241, (2009), K. Takiyama, E. Kinoshita, E. Kinoshita-Kikuta, Y. Fujioka, Y. Kubo, and T. Koike

Phos-tag beads as an immunoblotting enhancer for selective detection of phosphoproteins in cell lysatesAnalytical Biochemistry389, 83-85, (2009), E. Kinoshita-Kikuta, E. Kinoshita, and T. Koike

Mobility shift detection of phosphorylation on large proteins using a Phos-tag SDS-PAGE gel strengthened with agaroseProteomics9, 4098- 4101 (2009), E. Kinoshita, E. Kinoshita-Kikuta, H. Ujihara, and T. Koike

Separation and detection of large phosphoproteins using Phos-tag SDS-PAGENature Protocols4, 1513-1521 (2009), E. Kinoshita, E. Kinoshita-Kikuta, and T. Koike

A clean-up technology for the simultaneous determination of lysophosphatidic acid and sphingosine-1-phosphate by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using a phosphate-capture molecule, Phos-tagRapid Communications in Mass Spectrometry24, 1075-1084 (2010), J. Morishige, M. Urikura, H. Takagi, K. Hirano, T. Koike, T. Tanaka, and K. Satouchi

Genotyping and mapping assay of single-nucleotide polymorphisms in CYP3A5 using DNA-binding zinc(II) complexesClinical Biochemistry43, 302-306 (2010), E. Kinoshita, E. Kinoshita-Kikuta, H. Nakashima, and T. Koike

The DNA-binding activity of mouse DNA methyltransferase 1 is ragulated phosphorylation with casein kinase 1σ/εBiochemical Journal427, 489-497 (2010), Y. Sugiyama, N. Hatano, N. Sueyoshi, I. Suetake, S. Tajima, E. Kinoshita, E. Kinoshita-Kikuta, T. Koike, and I. Kameshita


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308-93563 Phos-tag™ Agarose
 Phos-tag 琼脂糖
3 mL