Immunostaining for Proliferating Cells

August 14, 2020

Vascular Dysfunction
University of Heidelberg
Postdoctoral Researcher

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Quality of Results

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Company:

Cell Signaling

Product Name:

Ki-67

Catalog Number:

9449

Clone Number:

8D5

Mouse embryonic brain cortex at E13 were stained with Ki-67 antibody to determine the population of proliferating cells

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

frozen embryonic mouse brain tissue (E13)

Primary Incubation

1:500, 4 degrees overnight

Blocking Agent

10% Normal donkey serum in PBST (0.5% Triton X 100)

Secondary Incubation

1:500, AF 488

Tertiary Incubation

none

Detection

Immunofluorescence

Results Summary

Although the progenitor cells were labelled with the antibody. The staining was not very clear. Some cells showed intense staining while others showed a faded. Should be used with another proliferation marker like PCNA.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Works if standardized properly.

The Bad

Antigen retrieval may be necessary for better staining, concentrated antibody may give unspecific signals.

The Bottom Line

Can be more commonly used for IHC, but not highly recommended for IF in tissues. Should be used with another cell proliferation marker for validation of results.

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