Cell lysates were equalized to protein content determined by Coomassie Plus™ Protein Assay Reagent (ThermoScientific #1856210) and loaded onto 4-12% NuPAGE Bis-Tris gels with MOPS running buffer with LDS Sample buffer supplemented with DTT. Gel proteins were transferred to PVDF membranes with an iBlot® Gel Transfer Device. 1X Casein-blocked membranes were probed with primary antibodies overnight at 4°C on an end-over-end rotisserie. Membranes were washed with TBS-T and HRP-conjugated secondary antibodies were added for 1 hour at room temperature. After washing, HRP was detected using Luminata™ Forte Western HRP Substrate reagent and recorded with a Bio-Rad VersaDoc imager.
Forte western hrp substrate reagent
Manufactured by Bio-Rad
The Forte Western HRP Substrate reagent is a chemiluminescent detection solution designed for the visualization of proteins in Western blot analysis. The reagent generates a luminescent signal upon reaction with the horseradish peroxidase (HRP) enzyme, which is commonly used as a reporter label in Western blotting applications.
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Lab products found in correlation
4 protocols using forte western hrp substrate reagent
1
Protein Immunoblotting for Quantitative Analysis
2
EGFR Protein Degradation Assay
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Example 41
Cell lysates were equalized to protein content determined by Coomassie Plus™ Protein Assay Reagent (ThermoScientific #1856210) and loaded onto 4-12% NuPAGE Bis-Tris gels with MOPS running buffer with LDS Sample buffer (supplemented with DTT. Gel proteins were transferred to PVDF membranes with an iBlot® Gel Transfer Device. 1× Casein-blocked membranes were probed with primary antibodies overnight at 4° C. on an end-over-end rotisserie. Membranes were washed with TBS-T and HRP-conjugated secondary antibodies were added for 1 hour at room temperature. After washing, HRP was detected using Luminata™ Forte Western HRP Substrate reagent and recorded with a Bio-Rad VersaDoc imager.
EGFR protein degradation was assessed by western blotting after treatment of T790M/L858R mutant Ba/F3 cell lines with a compound of the present application dose-dependently for 8 hour or in combination with 1 ug/mL of cetuximab. The results are shown in
3
Protein Degradation Analysis by Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 41
Cell lysates were equalized to protein content determined by Coomassie Plus™ Protein Assay Reagent (ThermoScientific #1856210) and loaded onto 4-12% NuPAGE Bis-Tris gels with MOPS running buffer with LDS Sample buffer (supplemented with DTT. Gel proteins were transferred to PVDF membranes with an iBlot® Gel Transfer Device. 1× Casein-blocked membranes were probed with primary antibodies overnight at 4° C. on an end-over-end rotisserie. Membranes were washed with TBS-T and HRP-conjugated secondary antibodies were added for 1 hour at room temperature. After washing, HRP was detected using Luminata™ Forte Western HRP Substrate reagent and recorded with a Bio-Rad VersaDoc imager.
EGFR protein degradation was assessed by western blotting after treatment of T790M/L858R mutant Ba/F3 cell lines with a compound of the present application dose-dependently for 8 hour or in combination with 1 ug/mL of cetuximab. The results are shown in
4
Protein Immunoblotting for Quantitative Analysis
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