Great Ready-to-Use Kit to Detect Cytokine Expression

Johns Hopkins School of Medicine
Oncology
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

BD Biosciences

Product Name:

Fixation/Permeabilization Solution Kit

Catalog Number:

554714

We routinely assess cytokine expression in T cells and macrophages using flow cytometry. Upon stimulation in the presence of a golgi inhibitor, we are able to prevent secretion of cytokines, allowing us to capture cytokine levels using flow antibodies to cytokines of interest. The kit fixes and perms the cells to allow intracellular detection.

Experimental Design and Results Summary

Application

Flow Cytometry

Starting Material

Single Cell Suspension of T cells or macrophages

Protocol Overview

Stimulate cells then surface stain cells before using 100 µL of ready-to-use 1x cytofix/perm kit (brown bottle) for 30 minutes at room temperature in the dark. Wash away the fix/perm using 1x wash buffer to then conduct intracellular staining using wash buffer.

Tips

Always use wash buffer to keep pores open in cells.

Results Summary

We are able to detect cytokine expression in cells using the kit and appropriate antibodies via flow analyzer.

DOI or PMID #

N/A

Additional Notes

Do not over fix since some cytokines are more sensitive to prolong fixation.

Image Gallery

Summary

The Good

Easy-to-use 1x solution and quite reliable every time for certain cytokines.

The Bad

None

The Bottom Line

Great cyto/fix perm kit to analyze cytokine expression. Very consistent results as long as you follow the same protocol.

Join the discussion