Great Resolution Even on Tumor Samples

April 10, 2019

Immunology
Columbia University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

FITC anti-mouse/human CD44

Catalog Number:

103006

Clone Number:

The goal of this study was to look at differences in the composition of memory, effector, and naiive T cell populations in tumor bearing mice. This antibody was used to help differentiate between memory and effector T cells that infiltrate the tumor as well as the surrounding draining lymph nodes.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Murine Prostate Tumor

Primary Incubation

1:200 for 20 mins at RT

Blocking Agent

N/A

Secondary Incubation

Fix/Perm at RT for 40 mins

Tertiary Incubation

ICS at RT for 30 mins

Detection

Flow Cytometry

Results Summary

Tumors were harvested and dissociated, and then stained for flow cytometry. The cells were first stained for extracellular markers, including CD4, CD8, CD62L, and CD44. Cells were then fixed and permeabilized for intracellular staining afterwards. This antibody offered great resolution to separate out CD44 and CD62L expressing CD8 T cells from the tumor, which is what is shown in the figure. Despite having low numbers of cells, which is the case with prostate tumors, the staining was still robust enough to detect differences in expression and separate cells into different populations. This stain was perfectly compatible with the rest of the 12 color panel and withstood fixation without a glitch.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Easy to optimize and use

The Bad

A little less bright than other colors

The Bottom Line

This antibody is easy to use and optimize, and a taking a spleen can serve as the perfect control to ensure that this antibody is working. It does have to be used a little more concentrated than our previous CD44 in a different color which makes it a little less cost-effective, but this is a great antibody if you have the FITC channel free in your panel.

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