Protease Inhibitor Cocktail (EDTA-Free, 200X 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.
- 1. Quanxin Jiang, Ning Wang, et al. "Targeting hepatic ceruloplasmin mitigates nonalcoholic steatohepatitis by modulating bile acid metabolism." J Mol Cell Biol. 2023 Sep 28:mjad060. PMID: 37771074
- 2. Md Shahjalal Hossain Khan, Marleigh Hefner, et al. "E4orf1 improves adipose tissue-specific metabolic risk factors and indicators of cognition function in a mouse model of Alzheimer's disease." Nutrition & Diabetes, 2023 Aug 12, 13(1): 13. PMID: 37573386
- 3. Xiaoxian Wu, Chengzhi Yu, et al. "The structural mechanism for transcription activation by Caulobacter crescentus GcrA." Nucleic Acids Res. 2023 Feb 28;51(4):1960-1970. PMID: 36715319
- 4. Junwan Lu, Yujie Li, et al. "Ciclopirox targets cellular bioenergetics and activates ER stress to induce apoptosis in non-small cell lung cancer cells." Cell Commun Signal. 2022 Mar 24;20(1):37. PMID: 35331268
- 5. Shi Du, Jiaxian Qian, et al. "Tumor cell-derived exosomes deliver TIE2 protein to macrophages to promote angiogenesis in cervical cancer." Cancer Lett. 2022 Mar 31;529:168-179. PMID: 35007697
- 6. Kathryn M Spitler, Shwetha K Shetty, et al. "Chronic high-fat feeding and prolonged fasting in liver-specific ANGPTL4 knockout mice." Am J Physiol Endocrinol Metab. 2021 Oct 1;321(4):E464-E478. PMID: 34396783
- 7. Kathryn M Spitler, Shwetha K Shetty, et al. "Regulation of plasma triglyceride partitioning by adipose-derived ANGPTL4 in mice." Sci Rep. 2021 Apr 12;11(1):7873. PMID: 33846453
- 8. Maria C Bewley, Lovely Gautam, et al. "Molecular architecture and domain arrangement of the placental malaria protein VAR2CSA suggests a model for carbohydrate binding." J Biol Chem. 2020 Oct 29;jbc.RA120.014676. PMID: 33122198
- 9. Liu W, Qin F, et al. "Sodium aescinate significantly suppress postoperative peritoneal adhesion by inhibiting the RhoA/ROCK signaling pathway." Phytomedicine. 2020;69:153193. PMID: 32120245
- 10. Monteiro-Brás T, Wesolowski J, et al. "Depletion of SNAP-23 and Syntaxin 4 alters lipid droplet homeostasis during Chlamydia infection." Microb Cell. 2019 Dec 3;7(2):46-58. PMID: 32025513
- 11. Wang H, Wang Z, et al. "Homotypic targeting upconversion nano-reactor for cascade cancer starvation and deep-tissue phototherapy." Biomaterials. 2020;235:119765. PMID: 31991338
- 12. Cap KC, Kim JG, et al. "P-Tyr42 RhoA GTPase amplifies superoxide formation through p47phox, phosphorylated by ROCK." Biochem Biophys Res Commun. 2020;523(4):972–978. PMID: 31973815
- 13. Zhang H, Read C, et al. "The Human Cytomegalovirus Nonstructural Glycoprotein UL148 Reorganizes the Endoplasmic Reticulum." mBio. 2019 Dec 10;10(6). pii: e02110-19. PMID: 31822584
在体外从培养基中提取分泌的蛋白质时,蛋白质产生停止,降解增加。细胞粗提取物中含有许多能够降解提取物中蛋白质的内源性酶,如磷酸酶和蛋白酶。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。
Protease Inhibitor Cocktail用于组织培养基以提高蛋白质的稳定性。该cocktail起到抑制降解非磷酸化或磷酸化蛋白质底物的蛋白酶的作用。这个cocktail用作组织培养基的补充物,以阻止分泌蛋白的降解。
该Protease Inhibitor Cocktail包含单独的组分,包括Aprotinin、Bestatin、E-64、Leupeptin和Pepstatin A,这些物质对半胱氨酸、丝氨酸、天冬氨酸和氨基肽酶具有广谱特异的抑制性。已经用组织培养基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溶剂为DMSO,作为即用型溶液提供。
建议适当地稀释cocktail以用于不同的细胞系。稀释测试至少以200倍的稀释倍数开始,因为DMSO浓度高于0.5%可能对细胞有害。由于不同的细胞系敏感性可能有差异,该cocktail可能需要进一步的稀释。这个cocktail的作用在培养基中可维持48小时,48小时后需要更换含有cocktail的新鲜培养基。
应用:WB,Co-IP,pull-down,IF,IHC,kinase assay等。
Catalog No. | Product Name | Summary | Targets | CAS Number | Smiles |
A2574 | Aprotinin | 牛胰蛋白酶抑制剂 | Proteases|Serine Protease | 9087-70-1 | CC(O)=O.CC.CC.CCccC.[R].[P].[2H].[L].[E].[P].[P].[Y].[3H].[G].[KH].[R].[R].[Y].F.[Y].[*].[L].F.[V].[Y].[G].[G].[R].[KH].[R].N.N.F.[KH].S.[*].[2H].[R].[G].[G].[*].[KH].[*].[3H].C.[*].F.[P].[*].I.I.N.[Q].[3H].[M] |
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, Microbial | 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 | 3-hydroxy-4-[2-[[3-hydroxy-6-methyl-4-[[3-methyl-2-[[3-methyl-2-(3-methylbutanoylamino)butanoyl]amino]butanoyl]amino]heptanoyl]amino]propanoylamino]-6-methylheptanoic acid |
-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。