The mKCs and normal human epidermal keratinocytes were lysed in radioimmunoprecipitation assay (i.e., RIPA) buffer after treatment, and total protein was quantified for each sample via bicinchoninic acid assay. Next, 5 μg of total protein was loaded onto a 4–20% Mini-PROTEAN TGX gel (Bio-Rad), transferred to a polyvinylidene difluoride membrane, and probed with primary antibodies against phosphorylated p38 (Cell Signaling, Danvers, MA; 1:100, rabbit), total p38 (Cell Signaling; 1:100, rabbit), cleaved caspase 3 (Cell Signaling; 1:500, rabbit), and phosphorylated-γH2AX (Abcam; 1:2,000, rabbit). Glyceraldehyde-3-phosphate dehydrogenase antibody (Abcam; 1:10,000, rabbit) was used as a loading control. Antirabbit secondary antibody conjugated to an infrared dye (IRDye800CW; Licor, Lincoln, NE) was used, and images were acquired on an Odyssey CLx imaging station (Licor).
Glyceraldehyde 3 phosphate dehydrogenase antibody
Manufactured by Abcam
Sourced in United States
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody is a protein that catalyzes the oxidative phosphorylation of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate in the glycolytic pathway.
Automatically generated - may contain errors
Lab products found in correlation
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Total p38, by Cell Signaling Technology (1 mentions)
Phosphorylated p38, by Cell Signaling Technology (1 mentions)
Irdye 800cw, by LI COR (1 mentions)
Mini protean tgx gel, by Bio-Rad (1 mentions)
Odyssey clx imaging station, by LI COR (1 mentions)
Imagequant las 4000, by GE Healthcare (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Clarity western ecl substrate, by Bio-Rad (1 mentions)
Hcv ns3 specific mouse monoclonal antibody clone h23, by Abcam (1 mentions)
Las 4000, by Cytiva (1 mentions)
2 protocols using glyceraldehyde 3 phosphate dehydrogenase antibody
1
Protein Profiling of Keratinocyte Stress Response
2
Western Blot Analysis of HCV NS3 Protein
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Treated Huh7it cells were lysed with radioimmunoprecipitation assay buffer and the amount of protein was calculated. Equal amounts of protein were separated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels and transferred onto polyvinylidene difluoride membrane (Millipore, Bedford, MA, USA). First antibody HCV NS3-specific mouse monoclonal antibody (clone H23; Abcam, Cambridge, MA, USA) and glyceraldehyde-3-phosphate dehydrogenase antibody (MBL) were incubated for 1 h, and phosphate-buffered saline-0.05% Tween was used for membrane washing. Second antibody horseradish peroxidase-conjugated goat anti-mouse immunoglobulin (MBL) was incubated for 1 h and the respective protein was visualized using Clarity Western ECL substrate (Bio-Rad). Skim milk (5%) was added to block the nonspecific binding for 60 min incubation. Chemiluminescence was detected using ImageQuant LAS 4000 (GE Healthcare).
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