Expression Of CD27 On Human PBMCs

December 21, 2016

Surgery
George Washington University, Washington DC
Post-doctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

APC/Cy7 anti-human CD27 Antibody

Catalog Number:

302816

CD27 is a co-stimulatory molecule expressed on T, B and NK cell surface and is a member of TNF- receptor superfamily. The CD27 marker in T cells divides the memory population into two groups, namely effector memory lacking CD27 and naive to transient memory which express CD27 on their surface. Thus expression of CD27 on CD4 T cells differentiates them into a non-effector type memory T cell which can be further classified by other surface markers such as CD62L , CCR7 and CD45RA expression.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMC

Primary Incubation

Human PBMC were stained with surface antibodies for CD3/CD4/CD27. Antibodies used were FITC anti-CD4, PE anti-CD3, and APC cy7 anti-CD27. Cells were stained for 30 mins in dark with antibody dilution of 1:100 for all flurochrome. Cells were washed twice with staining buffer after incubation and re-suspended in 500ul of staining buffer and acquired in 6 color flow cytometry. Analysis was done in a manner that CD3 T cells were gate first and CD4/CD27 expression was analyzed on that gate.

Blocking Agent

NA

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Flow cytometric detection

Results Summary

T cells that were gated by the expression of CD3 had a distinct population of CD4+ CD27+ T cells which was further sub-gated. comparison of this population across samples was important in determining the proportion of naive-transient to effector CD4 T cell population.

DOI or PMID #

NA

Additional Notes

Other surface markers like CD62L CCR7 and CD45RA may be helpful in determining memory vs naive vs effector cell population.

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Summary

The Good

Bright staining with APC/Cy7 anti-human CD27 Antibody was achieved which was helpful in distinguishing different cell subsets.

The Bad

Nothing to mention

The Bottom Line

Good quality surface antibody which requires minimal optimization and gives reliable results by flow cytometry.

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