Does Not Work for Mouse

November 01, 2019

BlueskyReddit
Medicine
UCLA
Postdoctoral Researcher

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Company:

BD Biosciences

Product Name:

Alexa Fluor® 647 Anti-Smad2 (pS465/pS467)/Smad3 (pS423/pS425)

Catalog Number:

562696

Clone Number:

O72-670

Browse Similar Products: SMAD2/SMAD3 Flow Cytometry Antibodies

While I have had great success with BD antibodies in the past, I regret to say that this antibody does not work for our purposes. On the product sheet, it says that it has been tested to react with mouse. However, I have been unable to get any positive phospho-SMAD signaling using flow cytometry.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse bone marrow cells

Primary Incubation

1:50

Blocking Agent

FBS

Secondary Incubation

-

Tertiary Incubation

-

Detection

Flow cytometer - BD FACS CANTOII

Results Summary

We harvested mouse bone marrow cells from the femurs and tibias of C57BL/6N mice. Cells were stimulated with TGFB (50ng, 30 minutes) or an equivalent volume of PBS and then fixed using the BD Fixation/Permeabilization Solution Kit and stained with Anti-Phospho-SMAD2/3. We do not see induction of SMAD signaling based on phosphorylation, which makes me believe that this antibody does not accurately detect phospho-SMAD2/3 in mouse cells for detection with flow cytometry.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Comes in multiple colors

The Bad

Does not work on mouse samples

The Bottom Line

This antibody does not work for our purposes, which includes analysis of mouse cells. It may work for human, but unfortunately is not a useful antibody for detection of phospho-SMAD activity in mouse cells.

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