Cell lysates were prepared by RIPA buffer with protease inhibitor cocktail (Roche). The protein concentrations of the lysates were quantified using BCA assay (#23225, Pierce, ThermoScientific). The protein lysates were heated for 5 min at 95°C in 4x SDS sample loading buffer (BioRad) and loaded into 10% SDS-Page gels. Afterwards samples were transferred onto PVDF membranes and the blots were probed with primary antibody in 5% Blotto (Thermo Fisher): (1:3000 rabbit anti-human NKD2 polyclonal antibody, Invitrogen PA5-61979) for 2 hours, followed by incubation with secondary antibody for 1 hour after washing (1:5000 horseradish-peroxidase -HRP-conjugated anti rabbit, Vector Laboratories) and developed using Pierce™ ECL Western Blotting Substrate A and B. Mouse monoclonal anti-GAPDH antibody (NovusBiologicals NB300-320; 1:1000) followed by HRP conjugated anti-mouse secondary antibody (Vector laboratories) was used as a loading control.
Horseradish peroxidase hrp conjugated anti rabbit
Manufactured by Vector Laboratories
Horseradish-peroxidase (HRP)-conjugated anti rabbit is a secondary antibody that is labeled with the enzyme horseradish peroxidase. This antibody is designed to bind to primary rabbit antibodies and can be used in various immunoassay techniques.
Automatically generated - may contain errors
Lab products found in correlation
Blotto, by Thermo Fisher Scientific (2 mentions)
Nb300 320, by Novus Biologicals (2 mentions)
Sds sample loading buffer, by Bio-Rad (2 mentions)
Pierce ecl western blotting substrate a and b, by Novus Biologicals (2 mentions)
Anti mouse hrp conjugated secondary antibody, by Vector Laboratories (2 mentions)
Protease inhibitor cocktail, by Roche (2 mentions)
2 protocols using horseradish peroxidase hrp conjugated anti rabbit
1
Western Blot Analysis of NKD2 Protein
2
Western Blot Analysis of NKD2 Protein
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Cell lysates were prepared by RIPA buffer with protease inhibitor cocktail (Roche). The protein concentrations of the lysates were quantified using BCA assay (#23225, Pierce, ThermoScientific). The protein lysates were heated for 5 min at 95°C in 4x SDS sample loading buffer (BioRad) and loaded into 10% SDS-Page gels. Afterwards samples were transferred onto PVDF membranes and the blots were probed with primary antibody in 5% Blotto (Thermo Fisher): (1:3000 rabbit anti-human NKD2 polyclonal antibody, Invitrogen PA5-61979) for 2 hours, followed by incubation with secondary antibody for 1 hour after washing (1:5000 horseradish-peroxidase -HRP-conjugated anti rabbit, Vector Laboratories) and developed using Pierce™ ECL Western Blotting Substrate A and B. Mouse monoclonal anti-GAPDH antibody (NovusBiologicals NB300-320; 1:1000) followed by HRP conjugated anti-mouse secondary antibody (Vector laboratories) was used as a loading control.
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