Alexa Fluor® 647 Anti-Human CD294 Antibody

January 07, 2019

JvGCRC
Nottingham Trent University
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Alexa Fluor® 647 anti-human CD294 (CRTH2) Antibody

Catalog Number:

350104

This antibody has been used to optimise flow cytometry-based immunophenotyping of whole-blood samples. The results have not yet been published. Multi-colour antibody panels were designed to identify a number of lymphocyte populations and tested to ensure appropriate compensation was possible. CD294 was used in a nine-colour panel to help identify group 2 innate lymphoid cells (ILCs). It is easy to use and works well.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human whole blood

Primary Incubation

5 µL AF647-CD294 for 30 minutes at 4°C

Blocking Agent

FcR blocking reagent, human (Miltenyi Biotec, cat# 130-059-901)

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

Beckman Coulter Galios, Kaluza software

Results Summary

Staining allowed the distinction of CD294+ cells although the separation from negative cells could be clearer. Titration of the antibody indicated that optimal staining was achieved at the manufacturers suggested dilution. This antibody fit well into a nine-colour panel with no compensation issues.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Staining of a rare population and useful for FACS analysis.

The Bad

The antibody has only been tested for flow cytometry and gives poor results if titrated.

The Bottom Line

Easy-to-use antibody pack that provided distinct staining of group 2 innate lymphoid cells population.

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