Human aoritic VSMCs were stimulated with TNFα (10 ng/mL, 10 min) then lysed (50mM Tris base, 150mM NaCl, 1mM EDTA, 10% glycerol, 1mM DTT, 1% Triton X-100, 0.1% Na-DOC, 0.1% SDS, 10mM β-glycerophosphate, 20mM para-nitrophenyl phosphate, 2mM sodium pyrophosphate, 1mM Na3VO4, 5mM NaF, 10 μg/ml aprotinin, and 1 mM phenylmethylsulfonyl fluoride (PMSF) at pH 7.4) for 1 hour with nutation at 4°C, and centrifuged for 30min at 20,000g. Supernatants were pre-cleared with protein-G sepharose beads for 1h at 4°C and cleared-supernatants were incubated with antibody (2 μg) for 1.5h, then incubated with protein-G sepharose for 1h. Beads were washed with lysis buffer, resuspended in SDS sample buffer, boiled and the associated proteins were then analyzed by western blot. Antibodies for IP were used as follows; Nox1 (sc-5821, Santa Cruz), LRRC8C (HPA029347, Sigma-Aldrich), LRRC8D (HPA014745, Sigma-Aldrich).
Nox1 sc 5821
Manufactured by Santa Cruz Biotechnology
Nox1 (sc-5821) is a primary antibody targeting the NADPH oxidase 1 (Nox1) protein. Nox1 is a subunit of the NADPH oxidase complex responsible for the production of superoxide radicals. This antibody can be used for the detection of Nox1 in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using nox1 sc 5821
1
TNFα-induced Signaling Pathway Analysis
2
TNFα-Induced Protein Interactions
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Cells were stimulated with TNFα (10 ng/mL, 10 min) then lysed (50mM Tris base, 150mM NaCl, 1mM EDTA, 10% glycerol, 1mM DTT, 1% Triton X-100, 0.1% Na-DOC, 0.1% SDS, 10mM β-glycerophosphate, 20mM para-nitrophenyl phosphate, 2mM sodium pyrophosphate, 1mM Na3VO4, 5mM NaF, 10 µg/ml aprotinin, and 1 mM phenylmethylsulfonyl fluoride (PMSF) at pH 7.4) for 1 hour with nutation at 4ºC, and centrifuged for 30min at 20,000g. Supernatants were pre-cleared with protein-G sepharose beads for 1h at 4ºC and cleared-supernatants were incubated with antibody (2 µg) for 1.5h, then incubated with protein-G sepharose for 1h. Beads were washed with lysis buffer, resuspended in SDS sample buffer, boiled and the associated proteins were then analyzed by western blot. Antibodies for IP were used as follows; Nox1 (sc-5821, Santa Cruz), LRRC8C (HPA029347, Sigma-Aldrich), LRRC8D (HPA014745, Sigma-Aldrich).
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