Specific and Stable Antibody for Detection of GFP

July 05, 2023

IKP/RBCT
Robert Bosch Gesellschaft für medizinische Forschung mbH
Graduate Student

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Quality of Results

Ease-of-Optimization

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Company:

Santa Cruz Biotechnology

Product Name:

GFP Antibody (B-2)

Catalog Number:

sc-9996

Clone Number:

B-2

This monoclonal IgG2a κ antibody from mouse works fine for the detection of GFP and some GFP-derived fluorophores in Western blot. Signals are specific and the antibody is stable at +4°C for a long time.

Experimental Design and Results Summary

Applications

Western blot

Sample

Human cancer cell lines (whole cell lysates) e.g. HEK293, MCF-7

Primary Incubation

1:1000 at 4°C overnight

Blocking Agent

5 % BSA in TBST

Secondary Incubation

1:2000, 60 min at RT

Tertiary Incubation

None

Detection

HRP, Chemiluminescence (West Dura Extended Duration Substrate) in Elysia-raytest detector (3 min exposure).

Results Summary

Detection is usually quite sensitive and shows bands at specific sizes for free EGFP (approx. 26 kDa) and EGFP-conjugated proteins (depending on the fused protein). No unspecific bands were observed in the cell lines analyzed (HEK293, MCF-7, DU145, Hela). Also, GFP derivate mTurquoise2 was successfully detected with this antibody, in contrast to mCitrine.

DOI or PMID #

N/A

Additional Notes

Diluted antibody solution can be stored at +4°C for at least two weeks with comparably good results. Stock solution can be stored for several years without noticeable quality loss. Dilution of 1:1000 works well in our hands but must possibly must be optimized depending on expression level. Other applications than Western blot are described by the supplier but were not tested.

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Summary

The Good

High sensitivity, fair price, binds (some) GFP derivates.

The Bad

Optimization for each GFP-fusion protein is recommended.

The Bottom Line

This GFP antibody from Santa Cruz Biotechnology is an easy-to-use Western blot antibody for the detection of any GFP-fusion protein or even other GFP derivates. We were happy with its sensitivity as well as long-term storage.

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