Total proteins were extracted, and equal amounts of protein lysate extracts were separated by 10% SDS-polyacrylamide gels and transferred to polyvinylidene difluoride membranes. Nonspecific binding was blocked with 5% skim milk for 2 hrs at room temperature. Membranes were incubated overnight at 4°C with primary antibodies for NLRP3 (Cell Signaling Technology), Caspase-1 (Cell Signaling Technology), ASC, ICAM-1 and VCAM-1 (Santa Cruz Biotechnology), washed and then incubated for 2 hrs at room temperature with horseradish peroxide-conjugated anti-rabbit or anti-mouse secondary antibodies (Santa Cruz Biotechnology). Blots were visualized with enhanced chemiluminescent substrate (Pierce Biotechnology). Relative quantities of proteins were determined with a densitometer.
Horseradish peroxide conjugated anti rabbit or anti mouse secondary antibodies
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Horseradish peroxidase-conjugated anti-rabbit or anti-mouse secondary antibodies are laboratory reagents used to detect and quantify target proteins in various immunoassay techniques. These antibodies are designed to bind to and label primary antibodies raised against rabbit or mouse antigens, enabling the visualization and measurement of the target proteins.
Automatically generated - may contain errors
Lab products found in correlation
Primary antibodies for nlrp3, by Cell Signaling Technology (1 mentions)
Vcam 1, by Santa Cruz Biotechnology (1 mentions)
Caspase 1, by Cell Signaling Technology (1 mentions)
Icam 1, by Santa Cruz Biotechnology (1 mentions)
Enhanced chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
2 protocols using horseradish peroxide conjugated anti rabbit or anti mouse secondary antibodies
1
Western Blot Analysis of NLRP3 Inflammasome
2
Western Blot Protein Analysis Protocol
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Proteins were extracted from cells, and lysates containing appropriate amounts of protein were resolved on 10% SDS-polyacrylamide gels and transferred to polyvinylidene difluoride membranes. Nonspecific binding was blocked in 5% bovine serum albumin for 2 h at room temperature. Membranes were incubated overnight at 4 °C with primary antibodies (Supplementary Table S2 ). Next, the membranes were washed and then incubated for 1 h at room temperature with horseradish peroxide-conjugated anti-rabbit or anti-mouse secondary antibodies (Santa Cruz Biotechnology, Dallas, TX, USA). The densitometric quantification of the protein bands was determined with the ImageJ software version 1.29x (National Institutes of Health, Bethesda, MD, USA).
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