Decent IFN-Gamma Antibody With High Sensitivity From BioLegend

August 26, 2024

BlueskyReddit
Cancer Center
George Washington University
PhD Candidate

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

APC anti-human IFN-γ Antibody

Catalog Number:

506510

Clone Number:

B27

In my research, I generate tumor-specific T cells and test their reactivity to tumor cell lines, including IFN-g secretion by T cells when co-cultured with tumor.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Human primary T cells

Primary Incubation

1:500-1:1000 viability stain in 100uL PBS, incubated at room temp for 15 mins

Blocking Agent

Human TruStain Fc block, 5uL diluted in 95uL FACS buffer (5% FBS in DPBS), incubated at 4C for 10 mins

Secondary Incubation

BD Cytofix/Cytoperm (follow manufacturer instructions)

Tertiary Incubation

1:100 anti-IFN-g antibody, incubate for 30 minutes at 4 degrees C

Detection

Flow cytometry in a BD Celesta or Cytek Aurora cytometer

Results Summary

This antibody has helped us discover which methods of generating tumor specific T cells result in highest tumor-specific activation, and has helped us further characterize which T cells in our product have tumor-specific activity.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

A little goes a long way.

The Bad

Using this product requires use of protein transport inhibitor and heavy optimization.

The Bottom Line

Antibody has great signal but requires a lot of titration and optimization for more complex flow panels.

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