Bm chemiluminescence western blot kit

Manufactured by Roche

Sourced in Germany

The BM chemiluminescence Western blot kit is a laboratory equipment product designed for Western blot analysis. It provides the necessary reagents and materials to detect and quantify target proteins using chemiluminescence detection.

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Lab products found in correlation

Pvdf membrane, by Merck Group (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) β actin antiserum, by Merck Group (1 mentions) Anti flag m2 peroxidase hrp antibody, by Merck Group (1 mentions)

4 protocols using bm chemiluminescence western blot kit

Evaluating prefrontal pNF-κB/NF-κB expression

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The changes in the expression of pNF-ĸB/NF-ĸB in the prefrontal cortex of the rat brain were evaluated by the Western blot assay. Following centrifuging at 12,000 rpm for 20 min at 4 °C, 10 mg of protein was determined on 10% SDS-PAGE gel and transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore, Germany). Five percent non-fat skim milk was used in order to obstruct membranes in 120 min. Then, membranes were incubated overnight with these primary antibodies: β-actin, NF-ĸB, and pNF-ĸB. All of the antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Tris-Buffered Saline with Tween 20 (TBST) was used for washing the membranes 3 times; then, they were incubated with secondary antibodies at room temperature for 1 h. The BM chemiluminescence Western blot kit (Roche, Mannheim, Germany) was used to detect bands, whereas the software ImageJ (Version 1.52t, Wayne Rasband (NIH), Bethesda, MD, USA) was used to evaluate their optimal density. The relative amount of pNF-ĸB/NF-ĸB was determined using GraphPad Prism 6 (San Diego, CA, USA).

Yousefi-Manesh H., Dehpour A.R., Shirooie S., Bagheri F., Farrokhi V., Mousavi S.E., Ricciutelli M., Cappellacci L., López V., Maggi F, & Petrelli R. (2021). Isofuranodiene, a Natural Sesquiterpene Isolated from Wild Celery (Smyrnium olusatrum L.), Protects Rats against Acute Ischemic Stroke. Pharmaceuticals, 14(4), 344.

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Quantification of Cholera Toxin and TCP

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Production of cholera toxin was determined by a ganglioside GM₁ enzyme-linked immunosorbent assay using a peroxidase-conjugated rabbit anti-cholera toxin B IgG (Pierce/Thermo Fisher Scientific) and a standard curve with varying concentrations of pure cholera toxin (Sigma Chemical Co.) as described previously [8 (link)]. TcpA, the major TCP subunit, was detected in western blots with a rabbit anti-TcpA serum. Cell pellets corresponding to 0.5 optical density units at 600 nm (OD₆₀₀) were boiled in 100 μL of sodium dodecyl sulfate — polyacrylamide gel electrophoresis loading buffer and proteins were separated in a 12 % polyacrylamide gel. Gels were transferred to polyvinylidene difluoride membranes and TcpA was detected by western blotting using a BM chemiluminescence western blot kit (Roche Applied Sciences, Indianapolis, IN). Flagellin FlaA expression was determined using the same procedure and the monoclonal antibody 3E1 raised against pure FlaA protein [10 (link)].

Wang H., Silva A.J, & Benitez J.A. (2017). 3-Amino 1,8-naphthalimide, a structural analog of the anti-cholera drug virstatin inhibits chemically-biased swimming and swarming motility in vibrios. Microbes and infection, 19(6), 370-375.

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Western Blot for NHE8 and β-catenin

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Total protein was prepared in a RIPA buffer. For Western blot detection, NHE8 antibody was used in a 1:2000 dilution to detect NHE8 protein,⁵ (link) β-catenin antiserum (catalog no. ab32572; Abcam, Cambridge, MA) was used in a 1:4000 dilution to detect β-catenin protein, and β-actin antiserum (A5316; Sigma) was used in a 1:5000 dilution to detect β-actin protein. Western detection was performed using the BM Chemiluminescence Western Blot Kit (catalog no. 11 520 709 001; Roche Diagnostics, Indianapolis, IN).

Xu H., Li J., Chen H, & Ghishan F.K. (2018). NHE8 Deficiency Promotes Colitis-Associated Cancer in Mice via Expansion of Lgr5-Expressing Cells. Cellular and Molecular Gastroenterology and Hepatology, 7(1), 19-31.

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Flag-tagged Protein Detection in Embryos

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Proteins were extracted from 80 injected embryos (24 hpf). Samples were dissolved in Laemmli buffer before gel electrophoresis. A mouse monoclonal ANTI-FLAG^® M2-Peroxidase (HRP) antibody (Sigma A8592) was used for detecting Flag-tagged protein using the BM chemiluminescence Western blot kit (Roche).

Joris M., Schloesser M., Baurain D., Hanikenne M., Muller M, & Motte P. (2017). Number of inadvertent RNA targets for morpholino knockdown in Danio rerio is largely underestimated: evidence from the study of Ser/Arg-rich splicing factors. Nucleic Acids Research, 45(16), 9547-9557.

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