Phosphatase Inhibitor Cocktail (2 Tubes, 100X)
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. Wei-Zhang S, Cui B, et al. "Chimpanzee adenovirus-mediated multiple gene therapy for age-related macular degeneration." Iscience, 2023. PMID: 37810255
- 2. Guo H, Peng X, et al. "Promoting Stem Cell Mechanosensing and Osteogenesis by Hybrid Soft Fibers." ACS Applied Materials & Interfaces, 2023. PMID: 37788009
- 3. Yuan-Li Ai, Wei-Jia Wang, et al. "Mannose antagonizes GSDME-mediated pyroptosis through AMPK activated by metabolite GlcNAc-6P." Cell Res. 2023 Jul 17. PMID: 37460805
- 4. Zhao B, Qiao H, et al. "HBV precore G1896A mutation promotes growth of hepatocellular carcinoma cells by activating ERK/MAPK pathway." Virol Sin. 2023 Oct;38(5):680-689. PMID: 37331658
- 5. Liu L, Yin S, Gan W. "TRAF6 Promotes PRMT5 Activity in a Ubiquitination-Dependent Manner." Cancers, 2023, 15(9): 2501. PMID: 37173967
- 6. Geng J, Kang Z, et al. "Microtubule Assists Actomyosin to Regulate Cell Nuclear Mechanics and Chromatin Accessibility." Research, 2023, 6: 0054. PMID: 37040508
- 7. Qiuhong Xiong, Huimin Sun, et al. "WDR45 mutation dysregulates iron homeostasis by promoting the chaperone-mediated autophagic degradation of ferritin heavy chain in an ER stress/p38 dependent mechanism." Free Radic Biol Med. 2023 May 20:201:89-97. PMID: 36940732
- 8. Zhang Q, Luo S, et al. "Upregulation of KHDC1L promotes the proliferation and inhibits apoptosis in head and neck squamous cell carcinoma." Epigenetics, 2023, 18(1): 2175168. PMID: 36734243
- 9. Sun Q, Pei F, et al. "Curved Nanofiber Network Induces Cellular Bridge Formation to Promote Stem Cell Mechanotransduction." Advanced Science, 2023, 10(3): 2204479. PMID: 36382560
- 10. Wei Y, Zhang D, et al. "PDK1 promotes breast cancer progression by enhancing the stability and transcriptional activity of HIF-1α." Genes & Diseases, 15 July 2023.
- 11. Zhu Z, Geng P, et al. "Highly Efficient Natural and Artificial Hybrid Photosynthesis Based on Carbon-Energy Coupling Transfer Enhancement by Formate with Engineering Chlamydomonas Reinhardtii Strains." Available at SSRN 4457571.
- 12. Gaojie Song, Jinbo Fang, et al. "Ad-apoptin inhibits glycolysis, migration and invasion in lung cancer cells targeting AMPK/mTOR signaling pathway." Exp Cell Res. 2021 Dec 15;409(2):112926. PMID: 34793774
- 13. Yi Fu, Yu Sun, et al. "Elevation of JAML promotes diabetic kidney disease by modulating podocyte lipid metabolism." Cell Metab. 2020 Dec 1;32(6):1052-1062.e8. PMID: 33186558
- 14. Yue P, Zhu C, et al. "Development of an autophagy-related signature in pancreatic adenocarcinoma." Biomed Pharmacother. 2020;126:110080. PMID: 32203889
- 15. 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
- 16. Wang J, Luo J, et al. "Increased intracellular Cl(-) concentration by activating FAK promotes airway epithelial BEAS-2B cells proliferation and wound healing." Arch Biochem Biophys. 2019 Dec 12:108225. PMID: 31838119
- 17. Reuven N, Adler J, et al. "Recruitment of DNA Repair MRN Complex by Intrinsically Disordered Protein Domain Fused to Cas9 Improves Efficiency of CRISPR-Mediated Genome Editing." Biomolecules. 2019 Oct 8;9(10). PMID: 31597252
- 18. DAyan A. Nurkesh, et al. "Date Pit Carbon Dots Induce Acidic Inhibition of Peroxidase and Disrupt DNA Repair in Antibacteria Resistance." Global challenges. 27 August 2019.
- 19. Cheng L, Li J, et al. "N-linoleyltyrosine protects against transient cerebral ischemia in gerbil via CB2 receptor involvement in PI3K/Akt signaling pathway. " Biol Pharm Bull. 2019 Sep 3. PMID: 31484847
- 20. Li T, Wei Z, et al. "Withanolides, Extracted from Datura Metel L. Inhibit Keratinocyte Proliferation and Imiquimod-Induced Psoriasis-Like Dermatitis via the STAT3/P38/ERK1/2 Pathway." Molecules. 2019 Jul 17;24(14). pii: E2596. PMID: 31319488
- 21. Reham Alsaadi. "Characterization of post-translational modification of ATG16L1 in antibacterial autophagy." University of Ottawa.2019.
蛋白磷酸化是重要的共价翻译后修饰，可以改变蛋白质的结构构象，并调控靶蛋白的功能，位置和特异性结合。许多细胞过程受蛋白质的可逆磷酸化调控，30％的蛋白质可能在其存在过程中被磷酸化。
由于内源性蛋白的产生和降解处于平衡状态，因此在稳定的环境条件下其细胞水平是稳定的。细胞粗提取物含有许多内源性酶，例如磷酸酶和蛋白酶，其能够降解和改变提取物中的蛋白。提高完整蛋白产量的最佳方法是加入这些酶的抑制剂。
更多的抑制剂意味着更好的磷酸化保护。我们提供两管具有不同溶液形式的抑制剂，帮助您更好地抑制磷酸酶。
管A（即Phosphatase Inhibitor Cocktail 1，货号K1012）抑制碱性磷酸酶的L-同功酶和丝氨酸/苏氨酸蛋白磷酸酶，例如蛋白磷酸酶1和2A（PP1和PP2A）。管A 含有Cantharidin、Bromotetramisole和Microcystin LR，以DMSO的即用溶液提供。
管B（即Phosphatase Inhibitor Cocktail 2，货号K1013）抑制酪氨酸蛋白磷酸酶，酸性磷酸酶和碱性磷酸酶。管B含有Sodium orthovanadate、Sodium molybdate、Sodium tartrate、Imidazole 和 Sodium Fluoride，以ddH2O的即用溶液提供。
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）依次加入A管和B管：加A管，混匀；再加B管，混匀（不可将A管和B管混合后再一起加入）。
WB，Co-IP，pull-down，IF，IHC，kinase assay等。
| Tube # | Catalog No. | Product Name | Summary | Targets | CAS Number | Smiles |
| A (100X in DMSO) | ||||||
| N1686 | Cantharidin | 蛋白磷酸酶1/2A抑制剂 | Natural Products | 56-25-7 | O=C([C@@]1(C([H])([H])[H])[C@]2(C([H])([H])[H])[C@@]3([H])O[C@]1([H])C([H])([H])C3([H])[H])OC2=O | |
| B4750 | (-)-p-Bromotetramisole Oxalate | ALP抑制剂 | Others|Others | 62284-79-1 | BrC(C=C1)=CC=C1[C@H]2N=C3SCCN3C2.OC(C(O)=O)=O | |
| B3698 | Microcystin-LR | 蛋白磷酸酶PP1/PP2A抑制剂 | Chromatin/Epigenetics|Protein Ser/Thr Phosphatases | 101043-37-2 | CC1C(NC(=O)C(NC(=O)C(C(NC(=O)C(NC(=O)C(NC(=O)C(=C)N(C(=O)CCC(NC1=O)C(=O)O)C)C)CC(C)C)C(=O)O)C)CCCN=C(N)N)C=CC(=CC(C)C(CC2=CC=CC=C2)OC)C | |
| B (100X in ddH2O) | ||||||
| A8524 | Sodium Orthovanadate | PTP抑制剂 | Membrane Transporter/Ion Channel|ATPase | 13721-39-6 | [O-][V](=O)([O-])[O-].[Na+].[Na+].[Na+] | |
| B7843 | Sodium molybdate | 酸性和磷蛋白磷酸酶抑制剂 | 7631-95-0 | [O-][Mo](=O)([O-])=O.[Na+].[Na+] | ||
| B7844 | Sodium tartrate | 酸性磷酸酶抑制剂 | 868-18-8 | C(C(C(=O)[O-])O)(C(=O)[O-])O.[Na+].[Na+] | ||
| B7845 | Imidazole | 碱性磷酸酶抑制剂 | 288-32-4 | C1=CN=CN1 | ||
| B7846 | sodium fluoride | 酸性磷酸酶抑制剂 | 7681-49-4 | [F-].[Na+] | ||
| -20°C条件下储存，稳定保存12个月以上。当在2-8°C条件下储存时，该产品可稳定保存2个月。 | ||||||
1）Q：可否将A、B两管产品混合后再加入裂解液中？
A：将A管和B管预先混合会形成浅绿色的沉淀，该沉淀主要是析出的盐类，沉淀会同时影响A管的盐类和B管的天然化合物。这些沉淀无法完全重新溶解。因此需要避免这种混合方式，而是按照说明书分别加入两管产品。
2）Q：Western Blot实验时磷酸化蛋白和非磷酸化蛋白的条带都非常弱应该怎么解决？
A：首先保证蛋白上样量足够；其次，需要同时加入蛋白酶抑制剂Cocktail（如货号K1007）和磷酸酶抑制剂Cocktail（如货号K1015）来保护蛋白不被降解。
3）Q：为什么A管的融化时间比B管的长？
A：因为A管的溶剂是DMSO，B管的溶剂是ddH2O，相比之下DMSO融化的速度更慢，特别是在低温条件下。可采用在25°C水浴中融化，亦可将A管分装成多个小管。
