Immunofluorescence of Ki-67 and Insulin in Paraffin Embedded Mouse Tissue

Molecular Physiology and Biophysics
Vanderbilt University Medical Center
Postdoctoral Research Fellow

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Quality of Results

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Company:

BD Pharmingen

Product Name:

Purified Mouse Anti-Ki-67

Catalog Number:

550609

We were determining the proliferative capacity of beta cells in a transgenic mouse model utilized in our lab. Ki-67 is a nuclear cell proliferation-associated antigen expressed in all active stages of the cell cycle, and was suitable for these purposes.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Mouse pancreatic paraffin-section

Primary Incubation

1:1000, 4 Deg C, overnight.

Blocking Agent

5% Normal Donkey Serum in 1%BSA/PBS (w/v)

Secondary Incubation

1:1000. Mouse IgG-Cy2. 2 hours, Room temperature

Tertiary Incubation

NA

Detection

Fluorescence microscopy

Results Summary

This antibody performed as expected. Several insulin positive beta cells were found proliferating as observed by immunofluorescence co-staining. See Yellow arrowheads in supplied image.

Additional Notes

Antigen retrieval (TEG, pH9.0) was necessary to detect Ki-67 in paraffin embedded sections.

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Summary

The Good

Excellent detection at 1:1000

The Bad

None

The Bottom Line

This antibody worked beautifully. Requires antigen retrieval if samples are embedded in paraffin.

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