Estrogen Receptor Antibody that Works Very Well for Mouse Xenografts

April 15, 2016

Center for Hearing & Deafness
University at Buffalo
Principal Investigator

Overall

Quality of Results

Ease-of-Optimization

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Company:

R & D Systems

Product Name:

Human ER alpha /NR3A1 Antibody

Catalog Number:

AF5715

We were studying tumor biomarkers in mouse xenografts by double immunofluorescent labeling experiments. We did not want to use mouse primary antibodies, due to excessive background issues and we already had a rabbit primary for another marker of interest. This Human ER alpha /NR3A1 Antibody from R& D Systems is made in sheep, which was ideal for our purposes.

Experimental Design and Results Summary

Applications

Immunohistochemistry

Sample

Paraffin section of mouse xenograft

Primary Incubation

AF5715 Human ER alpha /NR3A1 Antibody was used at 2 ug/ml in PBS-Tween for one hour at room temperature.

Blocking Agent

NA

Secondary Incubation

Life Technologies A21436 Donkey anti-Sheep Alexafluor 555 at 2 ug/ml in PBS-Tween for 45 minutes at room temperature

Tertiary Incubation

Vectashield mounting medium containing DAPI counterstain

Detection

Fluorescence microscopy- Nikon microscope with SPOT camera

Results Summary

In the attached image, you can see staining of the ER+ human tumor cells within the mouse xenograft. This was the first experiment and it did not need further troubleshooting. The ER alpha antibody showed good specificity with low background.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

Specific, low background, plays well with other primary antibodies.

The Bad

None

The Bottom Line

I would recommend this antibody for similar situations,--in the case of mouse xenografts where a mouse primary antibody is not ideal and a rabbit primary antibody is already part of the combination.

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