CD45RA with a Bright Conjugate (PE)

Toxicology
University of Kentucky
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Biosciences

Product Name:

PE Mouse Anti-Human CD45RA

Catalog Number:

555489

Clone Number:

Browse Similar Products: CD45RA Flow Cytometry Antibodies

The identification of naïve T cells relies on the correct characterization of T cells (using CD3), and then looking at their subsets (helper T cells: CD4+, and cytotoxic T cells: CD8+). These cells before they encounter an antigen (naïve T cells) will also express CD45RA. After antigen experience, central and effector memory T cells gain expression of CD45RO and lose expression of CD45RA. In our lab we are characterizing these subsets and so chose this antibody to go with the CD3/CD4/CD8/CD45RO panel we devised.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Peripheral Blood

Primary Incubation

15 minutes at 4 degrees on ice

Blocking Agent

BSA (1%)

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

In FL-2 channel (blue laser) flow cytometer

Results Summary

The majority of the lymphocytes shown in the plot (blue) stained positive with this antibody; however, we needed to do some optimization to get the signal right. Being so bright causes issues, especially with compensation.The plot on the left is the negative control.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Affordable. Bright fluorescence

The Bad

Too bright! Compensation issues inevitable

The Bottom Line

It's a good antibody, but you have to be careful with compensation, and it is imperative to titrate it.

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