Cells were lysed on ice, then 30 µg proteins were separated on 10% SDS-PAGE gels and transferred onto a polyvinyl difluoride (PVDF) membrane. After blocked with 5% skimmed milk, the membrane was incubated with one of the following antibodies overnight at 4 °C: anti-ASK-1 antibody (8662, Cell Signaling Technology), anti-phospho-ASK-1 antibody (3764, Cell Signaling Technology), anti-STRAP antibody (AP29336; One World Lab), anti-14-3-3 antibody (9636; Cell Signaling Technology), anti-caspase 8 antibody (ab25901, Abcam), anti-TNF-α antibody (ab1793, Abcam), and anti-tubulin antibody (ab6160, Abcam). Then the membrane was incubated with secondary antibody (ab6721 or ab6789, Abcam) for 1 h at room temperature. Finally, the membrane was photographed by ECL Imaging System (Tanon 6060, USA).
Anti ask 1 antibody
Manufactured by Cell Signaling Technology
Sourced in China
Anti-ASK-1 antibody is a research tool used to detect and study the apoptosis signal-regulating kinase 1 (ASK1) protein. ASK1 is a serine/threonine protein kinase involved in various cellular processes, including stress response and apoptosis. This antibody can be used for applications such as western blotting, immunoprecipitation, and immunohistochemistry to aid in the investigation of ASK1 expression and signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Ab6160, by Abcam (2 mentions)
Anti 14 3 3 antibody, by Cell Signaling Technology (2 mentions)
Ab1793, by Abcam (2 mentions)
Ab6721, by Abcam (1 mentions)
Ab25901, by Abcam (1 mentions)
Ab6789, by Abcam (1 mentions)
Ab32400, by Abcam (1 mentions)
Ecl plus kit, by Beyotime (1 mentions)
Ab178410, by Abcam (1 mentions)
Ripa lysate buffer, by Beyotime (1 mentions)
Ab2302, by Abcam (1 mentions)
3 protocols using anti ask 1 antibody
1
Western Blot Analysis of Apoptosis Signaling
2
Immunoprecipitation of ASK-1 Protein Complex
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Lysates were generated from cells. The lysates were precleared with Protein G beads. Thereafter, the precleared lysates were incubated with 5 µg/mL of anti-ASK-1 antibody (8662; Cell Signaling Technology) or normal rabbit IgG (5 µg/mL) using a constant rotation at 4 °C. Two hours later, protein G beads were added and incubated for another 2 h. After washed by lysis buffer, protein G beads were boiled in 1× Laemmli buffer, and the co-precipitated complexes were then separated on SDS-PAGE and used for western blotting analysis.
3
Western Blot Analysis of Cellular Signaling
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Cells were lysed with radioimmunoprecipitation assay (RIPA) lysate buffer (Beyotime, China). The same amount of protein was separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) gel and then transferred to a polyvinyl difluoride (PVDF) membrane. After soaking in protein-free fast block buffer (Beyotime, China), the membrane was incubated overnight with one of the following antibodies at 4°C: anti-ASK-1 antibody (8662, Cell Signaling Technology), anti-pASK-1 antibody (3764, Cell Signaling Technology), anti-STRAP antibody (AP29336; One World Lab), anti-14-3-3 antibody (9636; Cell Signaling Technology), anticleaved caspase-3 antibody (ab2302, Abcam), anti-TNF-α antibody (ab1793, Abcam), anti-MLL1 antibody (ab32400, Abcam), anti-WDR5 antibody (ab178410, Abcam), and anti-tubulin antibody (ab6160, Abcam). Then, the secondary antibody was used to incubate the membrane at room temperature for 0.5 h. Finally, the membrane was observed by ECL Plus Kit (Beyotime, China) and quantified using the ImageJ program (National Institutes of Health, USA).
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