Novus Biologicals
GFP Antibody
NB600-308
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G-protein coupled receptor kinase 2 (GRK2) phosphorylates and desensitizes activated GPCRs on the plasma membrane to regulate the GPCR on/off cycle in signal transduction events. GRK2 displays a diffuse cytoplasmic staining but upon GPCR activation gets recruited to the plasma membrane to then phosphorylate and down regulate activated GPCRs.In this experiment, GFP-tagged GRK2 was stably expressed in NIH-3T3 cells to determine the localization pattern. Since, the expression was very low, GFP antibody staining was done to amplify the signal. This GFP antibody was used based on my previous positive experience for staining other GFP tagged proteins.
Immunofluorescence
NIH-3T3 cells
1:500 dilution in blocking agent; 1 hr at room temp
1% Donkey serum + 1% BSA + 0.1% Triton-X 100 in 1xPBS
1:1000 dilution of Donkey anti-rabbit IgG-Alexa 488 conjugated
None
Confocal Microscopy
A diffuse cytoplasmic staining of GRK2-GFP was observed when expressed in NIH-3T3 cells
High signal and low noise
Good antibody for staining low expressed GFP tagged proteins