Protein blocking buffer (pH 7.5) (ab126587) for serum-free blocking of non-specific antibody binding in ICC, ELISA and western blot.
ELISA, Flow Cyt, WB, ICC/IF, In-Cell ELISA
Liquid
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
Application Flow Cyt | Reactivity No | Dilution info - | Notes - |
Application WB | Reactivity Reacts | Dilution info - | Notes - |
Application ICC/IF | Reactivity Reacts | Dilution info - | Notes - |
Application In-Cell ELISA | Reactivity Reacts | Dilution info - | Notes - |
Protein blocking buffer (pH 7.5) (ab126587) for serum-free blocking of non-specific antibody binding in ICC, ELISA and western blot.
ELISA, Flow Cyt, WB, ICC/IF, In-Cell ELISA
Liquid
Preservative: 100% Casein Blocking Buffer 10x
Ambient - Can Ship with Ice
Ambient
The product can be stored for up to 12 months
Protein blocking buffer (pH 7.5) (ab126587) for serum-free blocking of non-specific antibody binding in ICC, ELISA and western blot. BSA-free
Preparation: Dilute to 1X in deionized water or PBS (e.g. 1mL 10X blocking buffer + 9mL water/PBS). Diluting in PBS instead of water makes a somewhat more stringent blocking buffer due to the presence of salt in the PBS. Blocking Buffer is pH 7.5. The user should determine empirically whether dilution in water or PBS is most appropriate for the assay at hand. Use diluted block solution promptly.
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Primary :
All Lanes : top membranes with Anti AMPK pThr172 + Thr183 (Anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) antibody [EPR5683] ab133448) at 1/1000. Bottom membranes with Anti AMPKa (Anti-AMPK alpha 1 antibody [2B7] ab110036) at 1/1000
Lane 1 = AMPKa1 protein
Lane 2 = C2C12 cells treated with AICAR
Lane 3 = untreated C2C12 cells
Secondary: Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) and Goat polyclonal to Mouse IgG – H&L – Pre-Adsorbed (HRP) at 1:10,000 developed using the ECL technique.
Performed under reducing conditions (50mM DTT – Sample heated at 60°C). Predicted band size for AMPKa: 64 kDa Observed band size for AMPKa: 64 kDa
Left membrane All blocking steps = 1X blocking reagent ab126587 diluted in PBST
Right membrane All blocking steps = 5% Milk in PBST Exposure time : 5 minutes
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