Excellent Antibody for a Quick Snapshot of Dividing Cells

May 03, 2019

Immunology
Columbia University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Alexa Fluor® 700 anti-human Ki-67 Antibody

Catalog Number:

350529

Clone Number:

For this experiment we wanted to see what effects hypoxia had on the proliferation of human cell lines treated with cobalt chloride, an inducer of HIF-1a which leads to hypoxia-induced transcription. Following administration of colbalt chloride, a large population of cells became KI-67 low, indicating that they were not longer undergoing active proliferation.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

DU145 Cell Line

Primary Incubation

All ECS 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 1:50

Detection

Flow Cytometry

Results Summary

DU145 cells were incubated either with or without cobalt chloride to induce hypoxia. Cells were stained for markers of cell death as well as Ki-67. The Ki-67 antibody was used at 1:50 in a stain volume of 50 uL as part of the intracellular staining protocol. It compensated easily using beads. Based on the flow cytometry results, hypoxia reduces the population of cells that are proliferating at any given time. Plots shown are based on DU145 cells that were gated on live cells only using a fixable viability dye.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Easy to use and dinstinguishes cells clearly

The Bad

Difficult to find a really good negative control

The Bottom Line

Easy to use as part of intracellular staining protocol. Used at 1:50 dilution, so one vial can last for several assays and is a good value for the price. Bright staining that clearly separates populations.

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