Excellent Staining and Works Seamlessly with CD44

April 11, 2019

BlueskyReddit
Immunology
Columbia University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Brilliant Violet 510™ anti-mouse CD62L

Catalog Number:

104441

In this project, we aimed to quantify changes in the composition of tumor-infiltrating-lymphocytes in tumor bearing mice. This antibody was used alongside CD44 in order to separate populations of naive, effector, and memory CD8 T cells in the tumor.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Murine Prostate Tumor

Primary Incubation

1:400 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

Murine prostate tumors were mechanically dissociated and then plated for flow cytometry. This antibody was used at 1:400 as part of the primary extracellular stain. Afterwards, the cells were fixed and permeabilized and then stained for intracellular targets. This antibody held up well to fixation, and was stable for a week at 4C (which is how long the samples waited prior to running on the cytometer). The figure in this review shows a tumor draining lymph node from a tumor bearing mouse gated on Live CD45+ CD8+ CD4- cells. This antibody fits nicely into a multicolor flow panel and is moderately easy to compensate.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Bright staining, easy to delineate populations

The Bad

A bit difficult to compensate

The Bottom Line

This antibody is easy to use and optimize, and works well. A spleen or lymph node can be used as controls. Antibody can easily be fixed for running at a later date. Compensation can be a bit tricky when used in conjunction with other BV dies, especially BV605 and 650.

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