Good Antibody to Detect Proliferating and Cycling Cells

August 07, 2018

Oncology
Johns Hopkins School of Medicine
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Fisher (eBioscience)

Product Name:

Ki-67 Monoclonal Antibody (SolA15), FITC

Catalog Number:

11-5698-82

Our lab studies the role of mTOR signaling in CD8 T cell differentiation and function. T cell differentiation requires a dynamic change in transcriptional activity, protein translation, etc, and many of these events are tied to cell cycle. One key marker to identify highly proliferating cells is the use of Ki-67 as a marker. We routinely use this marker as a way to tell us of cycling T cells within our various experiments.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Murine Splenocytes

Primary Incubation

1:2000, 30 minutes, 4 deg in ebio Permwash buffer (Intracellular Staining)

Blocking Agent

FC block cells before surface panel staining

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

Flow Cytometer

Results Summary

We are easily able to analyze CD8 T cells undergoing cell proliferation versus cells that are not.

DOI or PMID #

N/A

Additional Notes

Please remember to fix and then permeabilize the nucleus with the transcription factor kit from eBio

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Summary

The Good

Very robust and bright antibody to identify cycling T cells.

The Bad

Must really titrate the antibody since it's quite bright.

The Bottom Line

Great antibody to mark Ki-67+ T cells. Also, the dilution is quite high for use, so overall fairly cheap since little antibody is required for staining.

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