Mouse Anti-CD4 Antibody For Use In Flow Cut-Metric Applications

September 22, 2017

Human Oncology
University of Wisconsin–Madison
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Tonbo Biosciences

Product Name:

FITC Anti-Mouse CD4 (GK1.5)

Catalog Number:

35-0041

This antibody is part of a 10 color flow panel to analyze and quantify different populations within tumor infiltrating lymphocytes. For this analysis we harvest tumors, create a single cell suspension and stain for the immune cell populations. The CD4 antibody is required to determine the T regulatory cell population and works very nicely together with the other antibodies in the panel. It gives a clear separation of positive and negative cells and facilitates the assessment of what percentage of CD4+ cells are also T regulatory cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Primary tumor tissue and spleen samples harvested from mice

Primary Incubation

1ul antibody per 3x10^6 cells in 100ul, incubated at RT for 40min

Blocking Agent

Staining was performed in PBS with 2% FBS

Secondary Incubation

Not necessary

Tertiary Incubation

Not necessary

Detection

It was detected with a flow cytometer (BD LSRFortessa)

Results Summary

The antibody gives a clear divide between positive and negative sample and makes selecting the positive population easy. Background staining is very minimal even though it is detected on the GFP channel. I have used this antibody for fluorescent staining of frozen sections but a different color conjugate yielded a lot better results.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Well-priced antibody, performs well, low background

The Bad

Have to use a little bit more of it than other antibodies in the same panel

The Bottom Line

Very good antibody, same quality as the same clone and color from other vendors but a lot cheaper.

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