Ki67 Immuno-fluorescence Staining

July 09, 2014

Pathology
Stanford
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Biolegend

Product Name:

Purified anti-mouse Ki-67 Antibody (clone 16A8)

Catalog Number:

652401

Our research lab studies immunology, in particular in cancer.

The antibody was used to examine tumor cell proliferation in the context of a spontaneous intestinal tumor model (the ApcMin mouse). The experiment was carried out to examine whether particular treatments would affect tumor cell proliferation in this model.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Fixed frozen mouse small intestine

Primary Incubation

1:100 in 1% BSA in PBS overnight at 4 degrees

Blocking Agent

5% goat serum in 1% BSA in PBS

Secondary Incubation

1:200 goat anti-rat total IgG Alexafluor 488

Tertiary Incubation

None

Detection

Zeiss confocal microscope

Results Summary

The antibody worked very well in detecting Ki67, and positive staining was very clear. (green is Ki67 staining and blue is DAPI)

Additional Notes

Ki67 stains the actively dividing stem cells in the intestinal crypts in the healthy surrounding tissue, stromal cells, and the actively dividing tumor cells (that extend beyond the crypts).

Related Categories

Image Gallery

Summary

The Good

Detection of the antigen was very clear and didn't require a lot of optimization to get it working.

The Bad

Nothing (the antibody was not tested for other methods like Western blotting)

The Bottom Line

The antibody is a great tool for detecting Ki67 using immunofluoresence.

Share your experience with other scientists. Write a Review! »

Join the discussion