Antioxidant peptide A

Antioxidant peptide A;

Antioxidant peptide A 是一种短肽。作用于癌细胞时,Antioxidant peptide A 的侧链有助于增强自由基清除活性。

Antioxidant peptide Aamp;;

Antioxidant peptide A Chemical Structure

规格 价格 是否有货
1 mg ¥900 询问价格 货期
5 mg ¥2700 询问价格 货期
10 mg ¥4400 询问价格 货期

* Please select Quantity before adding items.

Antioxidant peptide A 的其他形式现货产品:

Antioxidant peptide A TFA

生物活性

Antioxidant peptide A is a short peptide, which contains alternative aromatic or sulfur-containing amino acid. The side chains of Antioxidant peptide A are believed to contribute to strong radical scavenging activities of peptides in the cancer cell.

体外研究
(In Vitro)

The effects of 10-100 μM of Antioxidant peptide A (Pep-A) concentrations are studied on the superoxide dismutase (SOD) enzyme activity. The enzyme activity decreases by 0.5 and 0.7-folds at 10 and 50 µM Antioxidant peptide A concentrations, respectively, and increases by 1.79-folds at 100 µM Antioxidant peptide A treatment, indicating that this concentration can be ideal for the treatment on Y79 a, RB cells. Furthermore, the Antioxidant peptide A can be involved in decreasing the ROS by increasing the antioxidant enzyme activity. A similar increase in the antioxidative enzyme levels in the presence of Hoki skin antioxidative peptide in hepatocarcinoma cells is attributed to the peptide’s role in maintaining the redox balance in the cellular environment. Cell viability analysis results show that the Antioxidant peptide A shows no toxicity to cancerous (Y79) cells and non-cancerous cells even after 48 h of treatment. The Y79 RB cell viability ranges between 115 and 157 % and 111-126 % after 24 and 48 h of exposures with Antioxidant peptide A, respectively. The cancer cell death from the treatment of 10-100 µM GNPs concentration is studied[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

分子量

770.97

Formula

C31H54N12O7S2

Sequence

Pro-His-Cys-Lys-Arg-Met

Sequence Shortening

PHCKRM

运输条件

Room temperature in continental US; may vary elsewhere.

储存方式

Please store the product under the recommended conditions in the Certificate of Analysis.

Solvent Solubility
In Vitro:;

H2O

Peptide Solubility and Storage Guidelines:

1.;;Calculate the length of the peptide.

2.;;Calculate the overall charge of the entire peptide according to the following table:

; Contents Assign value
Acidic amino acid Asp (D), Glu (E), and the C-terminal -COOH. -1
Basic amino acid Arg (R), Lys (K), His (H), and the N-terminal -NH2 +1
Neutral amino acid Gly (G), Ala (A), Leu (L), Ile (I), Val (V), Cys (C), Met (M), Thr (T), Ser (S), Phe (F), Tyr (Y), Trp (W), Pro (P), Asn (N), Gln (Q) 0

3.;;Recommended solution:

Overall charge of peptide Details
Negative (lt;0) 1.;;Try to dissolve the peptide in water first.
2.;;If water fails, add NH4OH (lt;50 μL).
3.;;If the peptide still does not dissolve, add DMSO (50-100 μL) to solubilize the peptide.
Positive (gt;0) 1.;;Try to dissolve the peptide in water first.
2.;;If water fails, try dissolving the peptide in a 10%-30% acetic acid solution.
3.;;If the peptide still does not dissolve, try dissolving the peptide in a small amount of DMSO.
Zero (=0) 1.;;Try to dissolve the peptide in organic solvent (acetonitrile, methanol, etc.) first.
2.;;For very hydrophobic peptides, try dissolving the peptide in a small amount of DMSO, and then dilute the solution with water to the desired concentration.
参考文献
  • [1]. Kalmodia S, et al. Bio-conjugation of antioxidant peptide on surface-modified gold nanoparticles: a novel approach to enhance the radical scavenging property in cancer cell. Cancer Nanotechnol. 2016;7:1.

Kinase Assay
[1]

Y79 cells are seeded at 2×105 cells/well in 12-well plates with 1000 μL of culture media and incubated at 37°C overnight. The cells are then exposed to varying dosages of Antioxidant peptide A (Pep-A) and Pep-B (10, 50, and 100 μM) in fresh medium and then incubated for 6 h. At the end of the incubation, the cells are collected and washed with ice-cold PBS twice and lysed with cell lysis buffer [0.1 M Tris/HCl, pH 7.4 containing 0.5 % Triton X-100, 5 mM β-Mercaptoethanol, 0.1 mg/mL PMSF]. Cell lysate is centrifuged at 14000×g for 5 min at 4°C. The supernatant contains a combined SOD activity from cytosol and mitochondria. The SOD activity is measured using the superoxide dismutase (SOD) activity assay kit. In brief, the cell lysate, buffer, enzyme, and the WST reagent are diluted, and the solution is added as per the protocol and incubated at 37°C for 20 min and read at 450 nm on a microplate reader. SOD activity is then calculated as a percentage of the inhibition activity of xanthine oxidase[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Assay
[1]

Y79 retinoblastoma and MIOM1 cells are seeded at 5×103 cells/well in 96-well plates and incubated at 37°C overnight. The cells are then treated with varying concentrations (10, 30, 60, and 100 µM) of Antioxidant peptide A (Pep-A) and Pep-B in a fresh medium and incubated for specific time periods (24 and 48 h). At the end of the incubation, 10 µL of MTT (5 mg/mL) is added to the cells with fresh medium (100 µL) and incubated at 37°C until formazan crystals are formed. The formazan crystals are dissolved in 100 µL of DMSO, and the reading is taken at 570 nm. Cell viability is calculated[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

参考文献
  • [1]. Kalmodia S, et al. Bio-conjugation of antioxidant peptide on surface-modified gold nanoparticles: a novel approach to enhance the radical scavenging property in cancer cell. Cancer Nanotechnol. 2016;7:1.

日本同仁化学抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit| DOJINDO

上海金畔生物科技有限公司代理日本同仁化学 DOJINDO代理商全线产品,欢迎访问官网了解更多信息

抗氧化能力检测试剂盒 (DPPH法 )货号:D678
DPPH Antioxidant Assay Kit

DPPH Antioxidant Assay Kit
商品信息
储存条件:0-5度保存
运输条件:常温

特点:

● 即用型试剂盒、数据重现性好

● 配制试剂所需的时间大幅缩短

● 检测结果不易受pH, 溶剂等因素影响

下载说明书
产品文献

选择规格:
100tests
500tests

期货

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit
抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

规格性状
产品概述
操作时间大幅缩短
检测原理
检测步骤
检测结果不受各因素影响
检测例
参考文献
FAQ

规格性状

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

产品概述

近年来的研究发现,人体内的抗氧化能力的降低与各种疾病的产生息息相关,因此人们对于具有抗氧化能力的功能食品的需求越来越高。 根据日本高知大学岛村等人的文献报道 1),使用稳定的自由基 DPPH (2,2-Diphenyl-1-picrylhydrazyl)作为底物的抗氧化能力检测法是数据重现性最好的一种方法。本试剂盒依据岛村老师的检测方法,将DPPH法的过程进行了改良和标准化,解决了以往实验中孔间差较大、试剂配制过程繁冗等问题。

本试剂盒在日本高知大学农林海洋学部的岛村智子老师的指导下开发而成 。

1) T. Shimamura et al., Anal. Sci., 2014, 30, 717-721

操作时间大幅缩短

DPPH和Trolox在溶液状态下都不稳定,需要现配现用。特别是作为底物的DPPH,一般还需要检测吸光度来确定含量,因此操作的步骤和时间都十分冗长。本试剂盒已经将所需的试剂准备好并分成小份单独包装,检测前只需要溶解定容即可开始实验,大幅缩短了操作步骤和时间。(DPPH的溶解需要超声波振荡发生器)

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

检测原理

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

检测步骤

将试剂盒样品加至96孔板,培养30分钟后即可上机检测。

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

将试剂盒样品加至96孔板,培养30分钟后即可上机检测。

检测结果不受各因素影响

用DPPH法检测抗氧化能力的时候,溶液中的pH以及溶剂的浓度等因素会对检测结果产生影响。本试剂盒通过优化操作步骤、调整溶液添加顺序等方法最大程度抑制pH和溶剂浓度对检测结果的影响。

pH对检测结果的影响                                                样品溶剂对检测结果的影响

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

试剂盒内含的Assay Buffer可以保证检测反应在一定         样品量只占检测溶液体积的1/10(20 μl),因此样品的

的pH下进行。                                                                 溶剂无论是水还是无水乙醇,都不影响最终检测结果。

IC50值的复孔差

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

如果只用样品的抗氧化能力IC50值来评价,比较容易出现检测结果的波动。用标准物质(Trolox)和样品同时检测,通过Trolox等价活性值(TEAC)来评价的话,可以得到重现性高的检测结果。

TEAC( μg TE/μg)= Trolox IC50 (μg/ml)/ Sample IC50( μg/ml)

检测例

不同检测机构之间的检测结果的差异

下面3个不同的检测机构,使用本试剂盒检测已知的抗氧化物:没食子酸、儿茶酸、桑色素。用比色皿通过分光光度计检测Trolox等价活性值(TEAC),结果显示3个检测机构之间的检测结果基本上没有差异。

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

参考文献:T. Shimamura et al., NipponShokuhin Kagaku Kogaku Kaishi, 2007, 54, 482 – 487

分光光度计和酶标仪检测结果的一致性

按照上面的实验的同样的方法,改用酶标仪和96孔板进行检测并计算Trolox等价活性值(Trolox),检测结果也高度一致。

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

参考文献

1、Enjuro Harunari, Chiaki Imada, Yasuhiro Igarashi, “Konamycins A and B and Rubromycins CA1 and CA2, Aromatic Polyketides from the Tunicate-Derived Streptomyces hyaluromycini MB-PO13T”, J. Nat. Prod., 2019, 82, (6), 1609-1615.

2、Hiroki Ishida, Naoki Yamasaki, Yuuki Otsuka, Daichi Mori, Tomoko Shimamura, Takuya Hasegawa, Shuhei Ogo, Tadaharu Ueda, “Electrochemical Antioxidant Capacity Measurement: A Downsized System and Its Application to Agricultural Crops”, Anal. Sci., 2021, doi:10.2116/analsci.21P217.

3、M. A. Maky and T. Zendo, “Generation and Characterization of Novel Bioactive Peptides from Fish and Beef Hydrolysates”, Appl. Sci., 2021, doi:10.3390/app112110452.

4、S. Kato, K. Kuwata, “Pro-/anti-oxidative properties of dopamine on membrane lipid peroxidation upon X-ray irradiation”, Radiat. Phys. Chem., 2021, doi:10.1016/j.radphyschem.2021.109518.

5、F. F. Sofian, N. Kikuchi, T. Koseki, Y. Kanno, S. Uesugi, Y. Shiono, “Antioxidant p-terphenyl compound, isolated from edible mushroom, Boletopsisleucomelas”, Biosci., Biotechnol., Biochem., 2022, doi:10.1093/bbb/zbab224.

6、S. Jin, S. Kim, D. S. Kim, D. Son and M. Shin, “Optically Anisotropic Topical Hemostatic Coacervate for Naked-Eye Identification of Blood Coagulation”, Adv. Funct. Mater., 2022, doi:10.1002/adfm.202110320.

7、Mohamed Abdelfattah Maky,Takeshi Zendo,“Identification of a Novel Bioactive Peptide Derived from Frozen Chicken Breast Hydrolysate and the Utilization of Hydrolysates as Biopreservatives“,Bioactive Peptides in Health and Disease【A special issue of Biology (ISSN 2079-7737).】,2023,doi.org/10.3390/biology12091218

FAQ

Q:是否可以用涡旋振荡或者移液器吹打来溶解DPPH?
A:由于DPPH较难溶解,无法用涡旋振荡或者移液器吹打完全溶解。溶解不充分是造成误差的原因,请务必用超声振荡完全溶解。
Q:计算得到的IC50值的数据重现性不好,有哪些需要注意的地方?
A:(1)  请确认DPPH Reagent在溶解时,管内是否有残留。

由于DPPH较难溶解,请务必使用超声振荡器充分溶解后再使用。特别需要注意管底部是否有残留。具体的判断方法请参照下图或操作说明书。

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

(2) 样品的稀释浓度间隔过大,或者抑制曲线已经达到饱和的情况下检测出来的IC50值,可能会有较大差距。因此,请务必做预实验,摸索最佳浓度范围。

抗氧化能力检测试剂盒 (DPPH法 )货号:D678 DPPH Antioxidant Assay Kit

Q:每个试剂盒可以检测多少个样品?
A:<100 tests包装>

・每个试剂盒可以用于一块96孔板的检测

・为了确保数据的准确性,建议至少设置3个复孔,下面按照复孔数为3计算可检测的样品数。

・每个试剂盒的DPPH Reagent的量可以检测96孔板的100个孔。

(Blank 2和DPPH Reagent不需要添加,所以不计入孔数)

未知样品:可以检测1个样品

・对于未知的检测样品,需要做预实验,每个试剂盒可以检测1个样品。

<预实验的孔数:确认最佳浓度范围>

样品: 8个点(8个不同的梯度浓度系列) × 3 (3个复孔) = 24 well

Blank:1 (Blank 1) × 3 (3个复孔) = 3 well

<计算IC50所需要的孔数>

样品: 8个点(8个不同的梯度浓度系列) × 3 (3个复孔) = 24 well

Trolox:4个点(4个不同的梯度浓度稀释系列) × 3 (3个复孔) = 24 well

Blank:1 (Blank 1) × 3 (3个复孔) = 3 well

*不需要预实验时,可以多检测一个样品。

已知IC50值大致范围的样品:可以检测3个样品

・对于已知大致IC50值范围的样品,每个试剂盒可以检测3个样品。

对于不知道IC50值大致范围的未知样品,请按照操作步骤做预实验。

<计算IC50所需要的孔数>

样品: 8个点(8个不同的梯度浓度系列) × 3 (3个复孔) × 3 (3个样品)= 72 well

Trolox:4个点(4个不同的梯度浓度系列) × 3 (3个复孔) = 12 well

Blank:1 ( Blank 1) (n=3) = 3 well

 

<500 tests包装>

・每个试剂盒可以用于5块96孔板的检测(DPPH Reagent和Trolox Standard各5管)

・以下是500 tests包装可检测的样品数,100 tests包装请参考上面。

每个500 tests包装的试剂盒可以检测

(Blank 2和DPPH Reagent不需要添加,所以不计入孔数)

未知样品:可以检测8个样品

・对于未知的检测样品,需要做预实验,每个试剂盒可以检测8个样品。

<预实验的孔数:确认最佳浓度范围>

样品: 8个点(8个不同的梯度浓度系列) × 3 (3个复孔) × 8 (8个样品) = 192 well

Blank:1 (Blank 1) × 3 (3个复孔) × 3 (3块板) = 9 well

<计算IC50所需要的孔数>

样品: 8个点(8个不同的梯度浓度系列) × 3 (3个复孔) × 8 (8块板) = 192 well

Trolox:4个点(4个不同的梯度浓度稀释系列) × 3 (3个复孔) × 5 (5块板) = 60 well

Blank:1 (Blank 1) × 3 (3个复孔) × 3 (3块板) = 9 well

Q:如果从开始反应到检测的时间间隔比较长,是否会影响检测值?
A:如果反应时间过长,可能会影响检测结果。为了得到重现性高的检测数据,请严格按照操作说明书的步骤(25℃,30 min,避光)后,立即进行检测。检测多个孔板的时候,请保证各孔板的上机检测前的时间相同。
Q:食品样品如果进行样品的前处理?
A:关于食品样品的前处理方法,有下列检测实例供参考。<茶叶>

 

1)  取5 g 茶叶样品,加入50 ml 100℃的超纯水。

2)  在100℃下持续搅拌10 min。

3)10分钟搅拌后,用纱布过滤,并用超纯水将样品溶液调整至55 g。

4)将样品溶液转移至离心管,23℃,4000×g离心10 min。

5)用过滤膜(孔径0.45 μm)过滤上清液,制成样品溶液。

 

 

<青椒、红椒>

1) 将样品冻结干燥后用搅拌机打成粉末。

2) 取0.5 g粉末状的样品,添加2.5 g海砂和5 ml MWA提取溶剂。

(MWA溶剂的配制方法为 无水乙醇:超纯水:醋酸 = 90:9.5:0.5)

3) 搅拌10 s后,在超声波浴中37℃超声振荡5 min。

4) 23℃,1600×g离心10 min,回收上清。

5) 向沉淀物中再次加入5 ml MWA,重复步骤3,步骤4的操作3次。

6) 将所有回收得到的上清液加入25 ml容量瓶,用MWA定容作为检测样品。

Q:如果样品有混浊,是否可以检测?
A:<对于有混浊的样品>

样品中的混浊会影响检测结果。

我们尝试过用以下溶剂检测样品:水、无水乙醇、甲醇、WMA、DMSO。

*MWA溶剂的配制方法为 无水乙醇:超纯水:醋酸 = 90:9.5:0.5

*DMSO可能会造成TEAC值偏高。

如果样品不能完全溶解,仍然有混浊时,请将样品过滤后再使用。

混浊部分的抗氧化能力无法检测。

如果溶剂是水,还可以用SOD Assay Kit-WST检测。

由于DPPH的检测原理与SOD试剂盒的原理不同,可以用双指标进行验证。

另外,含有大量胡萝卜素的样品或者溶液呈紫色的样品不适用本试剂盒检测。

推荐用其他方法检测。

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Total Antioxidant Capacity (PAO) Test Kit

品牌:JaICA
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储存条件:+20°C
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Antioxidant peptide A TFA

Antioxidant peptide A TFA; 纯度: 99.35%

Antioxidant peptide A TFA 是一种短肽。作用于癌细胞时,Antioxidant peptide A 的侧链有助于增强自由基清除活性。

Antioxidant peptide A TFAamp;;

Antioxidant peptide A TFA Chemical Structure

规格 价格 是否有货 数量
1 mg ¥900 In-stock
5 mg ¥2700 In-stock
10 mg ¥4400 In-stock
50 mg ; 询价 ;
100 mg ; 询价 ;

* Please select Quantity before adding items.

Antioxidant peptide A TFA 相关产品

bull;相关化合物库:

  • Covalent Screening Library Plus
  • Bioactive Compound Library Plus
  • Anti-Cancer Compound Library
  • Covalent Screening Library
  • Peptide Library

生物活性

Antioxidant peptide A TFA is a short peptide, which contains alternative aromatic or sulfur-containing amino acid. The side chains of Antioxidant peptide A are believed to contribute to strong radical scavenging activities of peptides in the cancer cell[1].

分子量

884.99

Formula

C33H55F3N12O9S2

Sequence

Pro-His-Cys-Lys-Arg-Met

Sequence Shortening

PHCKRM

运输条件

Room temperature in continental US; may vary elsewhere.

储存方式

Sealed storage, away from moisture

Powder -80deg;C 2 years
-20deg;C 1 year

*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (sealed storage, away from moisture)

Solvent Solubility
In Vitro:;

H2O

Peptide Solubility and Storage Guidelines:

1.;;Calculate the length of the peptide.

2.;;Calculate the overall charge of the entire peptide according to the following table:

; Contents Assign value
Acidic amino acid Asp (D), Glu (E), and the C-terminal -COOH. -1
Basic amino acid Arg (R), Lys (K), His (H), and the N-terminal -NH2 +1
Neutral amino acid Gly (G), Ala (A), Leu (L), Ile (I), Val (V), Cys (C), Met (M), Thr (T), Ser (S), Phe (F), Tyr (Y), Trp (W), Pro (P), Asn (N), Gln (Q) 0

3.;;Recommended solution:

Overall charge of peptide Details
Negative (lt;0) 1.;;Try to dissolve the peptide in water first.
2.;;If water fails, add NH4OH (lt;50 μL).
3.;;If the peptide still does not dissolve, add DMSO (50-100 μL) to solubilize the peptide.
Positive (gt;0) 1.;;Try to dissolve the peptide in water first.
2.;;If water fails, try dissolving the peptide in a 10%-30% acetic acid solution.
3.;;If the peptide still does not dissolve, try dissolving the peptide in a small amount of DMSO.
Zero (=0) 1.;;Try to dissolve the peptide in organic solvent (acetonitrile, methanol, etc.) first.
2.;;For very hydrophobic peptides, try dissolving the peptide in a small amount of DMSO, and then dilute the solution with water to the desired concentration.
参考文献
  • [1]. Kalmodia S, et al. Bio-conjugation of antioxidant peptide on surface-modified gold nanoparticles: a novel approach to enhance the radical scavenging property in cancer cell. Cancer Nanotechnol. 2016;7:1.