Proteaseinhibitor cocktail 100x

Manufactured by Thermo Fisher Scientific

Sourced in United States

Proteaseinhibitor cocktail (100X) is a concentrated solution containing a mixture of protease inhibitors. It is designed to prevent the degradation of proteins by endogenous proteases in biological samples, enabling the preservation of protein integrity during extraction and analysis.

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Lab products found in correlation

Apolyvinylidene fluoride membrane, by Merck Group (1 mentions) T per tissue protein extraction reagent, by Thermo Fisher Scientific (1 mentions) Supersignal west pico chemiluminescent substrate, by Cytiva (1 mentions) Free glycerol reagent, by Merck Group (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Oleic acid, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Norepinephrine, by Merck Group (1 mentions) Protease k, by Merck Group (1 mentions) Hcs lipidtox green neutral lipid stain, by Thermo Fisher Scientific (1 mentions) Hcs lipidtox deep red neutral lipid stain, by Thermo Fisher Scientific (1 mentions) Bttaa, by Vector Laboratories (1 mentions) Trypsin inhibitor, by Worthington (1 mentions) Trypsin platinum, by Promega (1 mentions) Proteinase k, by Qiagen (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Ripa buffer, by Boston BioProducts (1 mentions) Anti zfx, by Cell Signaling Technology (1 mentions)

4 protocols using proteaseinhibitor cocktail 100x

Western Blot Protein Analysis Protocol

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Cells lysate was prepared using T-PER Tissue Protein Extraction Reagent (Thermo
Scientific) supplemented with Halt™ phosphatase and protease
inhibitor cocktail (100X) (Thermo Scientific). Proteins were separated on sodium
dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto a
polyvinylidene fluoride membrane (Millipore). After blocking with 5% skim
milk, the membrane was incubated with primary antibodies, followed by incubation
with horseradish peroxidase-conjugated secondary antibody. Target proteins were
visualized using Super Signal West Pico Chemiluminescent Substrate and
ImageQuant 800 (Amersham).

Lee R., Kang M.K., Kim Y.J., Yang B., Shim H., Kim S., Kim K., Yang C.M., Min B.G., Jung W.J., Lee E.C., Joo J.S., Park G., Cho W.K, & Kim H.P. (2021). CTCF-mediated chromatin looping provides a topological framework for the formation of phase-separated transcriptional condensates. Nucleic Acids Research, 50(1), 207-226.

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Comprehensive Lipid Analysis Protocol

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FFA fluorometric assay kit (Cayman, item no.700310); Lipofectamine 3000 (Invitrogen, L3000–015); Norepinephrine (Sigma, A9512); HCS LipidTOX^™ Deep Red Neutral Lipid Stain (Invitrogen^™, H34477); HCS LipidTOX^™ Green Neutral Lipid Stain (Invitrogen^™, H34477); Trypsin Platinum (Promega, VA9000); Protease K (Sigma, P2308); Oleic acid (Sigma, O1383); BSA (Sigma, A7030); FFA-free BSA (Millipore, Code82-002-4); Trypsin inhibitor (Worthington, LS003571); Protease inhibitor cocktail (100X) (Thermo scientific, 1861279); Free glycerol reagent (Sigma, F6428–40ML); Propargyl choline (Cayman, 25870); BTTAA (Click chemistry tools, 1236–100); CalFluor 647 Azide(Click chemistry tools, 1272–1); Phosphatidylcholine Assay Kit (Colorimetric/Fluorometric) (Abcam , ab83377); Phosphatidylethanolamine Assay Kit (Fluorometric) (Sigma, MAK361).

Zhang C., Ye M., Melikov K., Yang D., Dias do Vale G., McDonald J., Eckert K., Lin M.J, & Zeng X. (2024). CLSTN3B enhances adipocyte lipid droplet structure and function via endoplasmic reticulum contact. bioRxiv.

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Proteinase K Inactivation Protocol

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AAF/II fimbrial extract was incubated at 70°C for 10 min with 20 μg/mL Proteinase K (Qiagen) unless otherwise specified. In order to inactivate the enzyme, an EDTA-free protease inhibitor cocktail 100X (Thermo Scientific) was added to the extract and incubated for 15 min at 95°C. Proteinase K inactivation was corroborated by incubation of active and inactivated Proteinase K with BSA, followed by SDS-PAGE to evaluate protein integrity.

Alvestegui A., Olivares-Morales M., Muñoz E., Smith R., Nataro J.P., Ruiz-Perez F, & Farfan M.J. (2019). TLR4 Participates in the Inflammatory Response Induced by the AAF/II Fimbriae From Enteroaggregative Escherichia coli on Intestinal Epithelial Cells. Frontiers in Cellular and Infection Microbiology, 9, 143.

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Western Blot Analysis of ZFX in HeLa Cells

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At 72 h post-transfection, cellular proteins were extracted from HeLa cells with RIPA buffer (Boston BioProducts, Boston, MA, USA) containing 5 mM sodium fluoride (NaF), 1 mM sodium vanadate (Na₃VO₄), and 1 mM phenylmethylsulfonyl fluoride (PMSF), supplemented with 1% Protease Inhibitor Cocktail (100X, Thermo Scientific, Rockford, IL, USA). Protein concentration was measured with the Bradford Assay. 50 µg of protein was loaded and separated in an 8% polyacrylamide SDS-gel and electrotransferred onto polyvinylidene fluoride membranes. The membranes were incubated with primary antibodies, followed by incubation with matching HRP-conjugated secondary antibodies. Color development was carried out using the ECL reagents (Pierce, Rockford, IL, USA). GAPDH was detected and the results were used as protein loading controls. The primary antibodies used were: anti-ZFX (1:1000, Cell Signaling, Beverly, MA, USA) and anti-GAPDH (1:1000, Santa Cruz, TX, USA).

Xu S., Duan P., Li J., Senkowski T., Guo F., Chen H., Romero A., Cui Y., Liu J, & Jiang S.W. (2016). Zinc Finger and X-Linked Factor (ZFX) Binds to Human SET Transcript 2 Promoter and Transactivates SET Expression. International Journal of Molecular Sciences, 17(10), 1737.

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