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T3924

胰蛋白酶-EDTA 溶液

Name: 胰蛋白酶-EDTA 溶液
Brand: SIGMA

1 ×, sterile; sterile-filtered, BioReagent, suitable for cell culture, 0.5 g porcine trypsin and 0.2 g EDTA, 4Na per liter of Hanks′ Balanced Salt Solution with phenol red

别名:

Cell dissociation enzyme

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关于此项目

EC 号:

3.4.21.4 (BRENDA, IUBMB)

UNSPSC Code:

12352204

NACRES:

NA.78

MDL number:

MFCD00130286

Biological source:

Porcine

Concentration:

1 ×

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属性

biological source

Porcine

Quality Level

400

sterility

sterile; sterile-filtered

product line

BioReagent

form

solution

mol wt

23.4 kDa

concentration

1 ×

technique(s)

cell culture | mammalian: suitable, single cell analysis: suitable

impurities

Porcine parvovirus, none detected (9 CFR)

pH

7.0-8.0

shipped in

dry ice

storage temp.

−20°C

说明

Application

该产品的典型用途是使黏附细胞从培养表面脱落。使细胞从附着面脱落所需的胰蛋白酶浓度主要取决于细胞类型和培养阶段。

适合用于制备测序用单细胞悬液。

Biochem/physiol Actions

胰蛋白酶可切割赖氨酸和精氨酸残基的C末端肽段。如果酸性残基位于切割位点的任意一侧，则该反应的水解速度减慢，并且如果脯氨酸残基位于切割位点的羧基侧，则水解停止。胰蛋白酶活性的最佳pH条件为7-9。胰蛋白酶还可以切割氨基酸合成衍生物的酯和酰胺键。将EDTA添加到胰蛋白酶溶液中作为螯合剂，用于中和钙离子和镁离子以防止其抑制胰蛋白酶活性。去除这些离子会增加酶活性。

丝氨酸蛋白酶抑制剂（包括DFP、TLCK、APMSF、AEBSEF和抑肽酶等）会抑制胰蛋白酶。

Preparation Note

本产品含有酚红。由于在干冰上运输，包装中可能累积大量二氧化碳。这些CO₂可能会进入溶液并略微降低pH值，使溶液呈橙色而不是粉红色。橙色溶液仍然可以使用，或者可以用氢氧化钠调节pH。使用高浓度胰蛋白酶长时间孵育细胞，会破坏细胞膜并杀死细胞。溶解胰蛋白酶或稀释浓缩胰蛋白酶溶液时，应使用不含Ca²⁺ 或Mg²⁺的缓冲盐溶液。

Other Notes

胰蛋白酶是由含223个氨基酸残基的单链多肽组成的，它通过切割胰蛋白酶原的Lys-1le肽键以去除N-末端六肽而获得。氨基酸序列通过6个二硫键交联。这是天然形式的胰蛋白酶——β-胰蛋白酶。 β-胰蛋白酶可以自溶，在Lys-Ser残基处断裂，产生α-胰蛋白酶。胰蛋白酶是丝氨酸蛋白酶家族的成员。

Disclaimer

本品应置于-10至-40°C环境冷冻储存。避免反复冻融。

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存储类别

12 - Non Combustible Liquids

wgk

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

历史批次信息供参考:

分析证书(COA)

Lot/Batch Number

0000445328

0000406896

0000424253

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