Good Option to Stain for Ki67

December 15, 2022

Charite Strasse 1
Deutsches Rheuma-Forschungszentrum
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

PE/Dazzle™ 594 anti-mouse/human Ki-67 Antibody

Catalog Number:

151219

Clone Number:

11F6

Info:Supplier Page View Product Specs

Browse Similar Products: Ki-67 Antibodies

In different healthy and pathological contexts, CD4 and CD8 T cells differentiate into a range of specialized effector and regulatory subsets. In mouse models of aging and autoimmune diseases we study the proliferation of CD4 and CD8 T cells. We identified Ki671+ proliferating cells with an anti-Ki67 PE-Dazzle594 antibody in pre-lupus NXBxW F1 mice.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Splenocytes of a NZBxW mouse, incubated in complete culture medium at 37 °C with (Restimulated) or without (unstimulated) PMA/ionomycin for 4h and brefeldin A for the 3 last hours.

Primary Incubation

Washed twice in PBS and incubated with L/D fixable aqua dye 20 min at 4°C, then washed twice in PBS.

Blocking Agent

2.4G2

Secondary Incubation

Surface staining with anti-CD3 BUV805, etc., in Cell Staining Buffer (Biolegend) incubated 30 min at 4°C.

Tertiary Incubation

Cells were fixed and permeabilized with commercial kit (TrueNuclear Biolegend) for 45 min at 4°C with fixative, 15 min at 4°C with permeabilization buffer, and stained for cytokines and anti-Ki67 PE-Dazzle 594 1/600 in permeabilization buffer.

Detection

Yellow-green laser on Symphony cytometer

Results Summary

We could identify and quantify Ki67+ proliferating CD3+ T cells. PMA+Ionomycin increases the proportion of Ki67+ proliferating cells. There are different levels of Ki67 expression.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Bright, gives good separation at relatively high dilution.

The Bad

Some cells are dim. A FMO should be done to set the positive threshold.

The Bottom Line

Very good option, increase the size of a panel advantageously. Cheap enough, given the high dilution.

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