Not Very Bright IL2 Staining, but Usable

July 01, 2020

Charite Strasse 1
Deutsches Rheuma-Forschungszentrum
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Brilliant Violet 711™ anti-mouse IL-2 Antibody

Catalog Number:

503837

Clone Number:

JES6-5H4

In different healthy and pathological contexts CD4 T cells differentiate into a range of specialized interleukin-secreting subsets. In mouse models of aging and autoimmune diseases we study the plasticity and effector functions of CD4 T helper cells. We used anti-IL-2 BV711 to quantify the number of CD4 T cells expressing IL-2 in old C57Bl/6J mice.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Splenocytes of wild-type 12 months-old C57Bl/6J mouse, incubated in complete culture medium at 37 °C with PMA/ionomycin for 4h and brefeldin A for the last 3 hours.

Primary Incubation

Surface staining with anti-CD4 Al700 in PBS 0.5% BSA incubated 30min at 4°C. Washed twice in PBS and incubated with L/D fixable aqua dye 20 min at 4°C, then washed twice in PBS.

Blocking Agent

clone 2.4G2 (Fc block)

Secondary Incubation

Cells are finally incubated with BV711 anti-IL-2 at 1/100 and PE Cy7 anti-IFN-g antibody in permeabilization buffer for 30 min at 4°C and washed once with permeabilization buffer and once with PBS BSA 0.5%.

Tertiary Incubation

N/A

Detection

Flow cytometry, violet laser

Results Summary

I could quantify IL-2-producing CD44+ CD4 T memory cells in the spleen.

Image Gallery

Summary

The Good

The antibody works well but is not very bright compared to other conjugates

The Bad

Spillover of PerCP Cy5.5 into BV711 channel can be high

The Bottom Line

Good option to increase a panel, but not the brightest, and need to be carefully compensated.

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