Protease Inhibitor Cocktail(EDTA-Free,100X in DMSO)
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
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在平衡状态下内源性蛋白质产生并降解,因此在稳定的环境条件下其细胞水平是稳定的。粗细胞提取物含有许多能够降解提取物中的蛋白质的内源酶,例如磷酸酶和蛋白酶。当在体外从细胞和组织中提取蛋白质时,蛋白质产生被显著抑制,降解增加。提高完整蛋白质产量的最佳方法是加入已知存在的那些酶的抑制剂。
蛋白酶抑制剂cocktail用于细胞裂解物或组织提取物以提高蛋白质的稳定性。该cocktail起到抑制蛋白酶的作用,这种蛋白酶会降解非磷酸化或磷酸化蛋白质底物。
该蛋白酶抑制剂cocktail包含单个组分,包括对半胱氨酸,丝氨酸,酸性蛋白酶和氨基肽酶具有广泛特异性的AEBSF,Aprotinin,Bestatin,E-64,Leupeptin 和 Pepstatin A。已经用哺乳动物细胞和组织提取物对这种蛋白酶抑制剂cocktail进行了优化和测试。该蛋白酶抑制剂cocktail作为DMSO即用溶液提供。
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在室温下解冻,然后在实验前以1:100(v / v)稀释至溶液样品(如细胞裂解物或组织提取物)。
应用:WB,Co-IP,pull-down,IF,IHC,kinase assay等。
Catalog No. | Product Name | Summary | Targets | CAS Number | Smiles |
A2573 | AEBSF.HCl | 丝氨酸蛋白酶抑制剂 | Proteases|Serine Protease | 30827-99-7 | C1=CC(=CC=C1CCN)S(=O)(=O)F.Cl |
A2574 | Aprotinin | 牛胰蛋白酶抑制剂 | Proteases|Serine Protease | 9087-70-1 | |
A2575 | Bestatin | 氨肽酶抑制剂 | Proteases|Aminopeptidase | 58970-76-6 | CC(C)CC(C(=O)O)NC(=O)C(C(CC1=CC=CC=C1)N)O |
A2576 | E-64 | 半胱氨酸蛋白酶(cysteine protease)抑制剂 | Proteases|Cathepsin | 66701-25-5 | CC(C)CC(C(=O)NCCCCN=C(N)N)NC(=O)C1C(O1)C(=O)O |
A2570 | Leupeptin | 丝氨酸和半胱氨酸蛋白酶抑制剂 | Proteases|Serine Protease | 103476-89-7 | O=C(C(C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H])([H])N([H])C(C([H])([H])[H])=O)N([H])C(C(N([H])C(C([H])=O)([H])C([H])([H])C([H])([H])C([H])([H])/N=C(N([H])[H])/N([H])[H])=O)([H])C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H] |
A2571 | Pepstatin A | 天冬氨酸蛋白酶抑制剂 | Proteases|Other Proteases | 26305-03-3 | CC(C)CC(C(CC(=O)O)O)NC(=O)C(C)NC(=O)CC(C(CC(C)C)NC(=O)C(C(C)C)NC(=O)C(C(C)C)NC(=O)CC(C)C)O |
-20°C条件下储存,稳定12个月以上。 |
1)Q:实验中经常用PMSF来保护蛋白,蛋白酶抑制剂Cocktail和它比有什么优势?
A:PMSF只能抑制一种蛋白酶(丝氨酸蛋白酶),Cocktail抑制的更全面且更加持久;此外,PMSF毒性较高并且PMSF更容易降解失效。
2)Q:抑制剂Cocktail可以反复冻融吗?
A:可以反复冻融,但是尽量减少反复冻融的次数。
3)Q:应该选择片剂的Cocktail还是溶液型Cocktail?
A:两种类型的产品都含有添加物,片剂中含有若干种公司专利的未经公开的医药成分,而溶液型产品中含有溶剂成分DMSO。一般来说这些添加物是不会影响产品效果的。溶液型需要的加工步骤少,因而质量好价格低。而片剂产品在储存和运输两个环节具有优势。(片剂能够在4°C储存而不需要-20°C)。
4)Q:用了Cocktail,蛋白仍然降解是怎么回事?
A:蛋白酶抑制剂Cocktail作用是保护蛋白免受蛋白酶降解,提高蛋白得率。对于绝大多数蛋白,Cocktail都有很好的效果。如果还是出现蛋白得率低的情况,可检查实验操作步骤和设计,减少导致蛋白降解的环节。例如,细胞处理前,先要将Cocktail加入到裂解液中并混匀,收集完细胞后立即加入准备好的裂解液。