Mouse Anti-GFP Antibody for Western Blots

December 09, 2015

Biology
Franklin and Marshall College
Assistant Professor

Overall

Quality of Results

Ease-of-Optimization

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Company:

GeneTex

Product Name:

Mouse anti-GFP [GT859]

Catalog Number:

GTX628528

This antibody was on immunoblots from HeLa cells transfected with an GFP-containing constructs. GT589 showed moderate to strong immunoreactivity at the expected molecular weights. Background levels were acceptable, but not absent. This antibody can be used in place of JL-8 from Clontech which shows a higher level of background reactivity especially at the lower molecular weights

Experimental Design and Results Summary

Applications

Western Blot

Sample

HeLa cell lysates

Primary Incubation

GT859 1:5000 in 5% milk in PBS + 0.1% tween-20 for 90 minutes

Blocking Agent

5% milk in PBS + 0.1% tween-20

Secondary Incubation

Goat anti-mouse HRP (Santa Cruz Biotech)1:5000 in 5% milk in PBS + 0.1% tween-20 for 90 minute

Tertiary Incubation

NA

Detection

ECL visualized on a Protein Simple FluorChem Q

Results Summary

GT859 worked as well as JL-8 in similar experiments. Both antibodies have a considerable level of background staining.

Additional Notes

None

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Summary

The Good

Detects GFP containing constructs in western blot

The Bad

Price is expensive and background issues persist

The Bottom Line

If JL-8 gives background staining in your region of interest (low molecular weight) the GT859 might provide an alternative.

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