GFP Antibody Roche: Works Well

Biochemistry
Heidelberg University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Roche

Product Name:

GFP antibody clones 7.1 and 13.1

Catalog Number:

11814460001

Clone Number:

N/A

GFP antibody has been used to detect the GFP signal after the transfection of an empty-GFP plasmid in Hela cells.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Hela cells, cell lysate

Primary Incubation

1:2000 Milk 5% in PBS 0,05% tween 1h RT

Blocking Agent

Milk 5% in PBS 0,05% tween

Secondary Incubation

Mouse 1:3000 milk 5% in PBS 0,05% tween 45 minutes

Tertiary Incubation

None

Detection

ECL

Results Summary

Hela cells WCL has been loaded on a 4-12% SDS-precast gel (1:30h; 180 V) then proteins have been transferred on nitrocellulose membrane using the semi-dry blotting (1h 12V). Blocking with milk 5% in PBS 0,05% tween (45 minutes) was followed by incubation overnight at room temperature with anti- GFP antibody (1:2000 in milk 5% in PBS 0,05% tween). Then the membrane was washed with 3x 5 minutes with PBS 0,05% tween, followed by a 45 minutes incubation with Mouse secondary antibody 1:3000 in milk 5% in PBS 0,05% tween. The secondary antibody was washed out with a second wash with PBS 0,05% tween (3x 5 minutes). ECL substrate has been used to detect the signal.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Very good staining and results obtained.

The Bad

Sometimes the signal can be very strong, the optimization of the dilution of the antibody between 1:2000-5000 can helo

The Bottom Line

Great antibody

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