GFP Antibody For Immunofluorescence Staining

Biochemistry
Stanford University
Senior Research Scientist

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Quality of Results

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Company:

Novus Biologicals

Product Name:

GFP Antibody

Catalog Number:

NB600-308

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.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

NIH-3T3 cells

Primary Incubation

1:500 dilution in blocking agent; 1 hr at room temp

Blocking Agent

1% Donkey serum + 1% BSA + 0.1% Triton-X 100 in 1xPBS

Secondary Incubation

1:1000 dilution of Donkey anti-rabbit IgG-Alexa 488 conjugated

Tertiary Incubation

None

Detection

Confocal Microscopy

Results Summary

A diffuse cytoplasmic staining of GRK2-GFP was observed when expressed in NIH-3T3 cells

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

High signal and low noise

The Bad

None

The Bottom Line

Good antibody for staining low expressed GFP tagged proteins

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