Use Of Rat IgG1 Kappa Isotype Control - EFluor 450 As A Control During Flow Cytometric Analysis Of CD4+ T Cells

Pharmacology and Experimental Neuroscience
University of Nebraska Medical Center
Graduate Research Assistant

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Quality of Results

Ease-of-Optimization

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Company:

Invitrogen Antibodies

Product Name:

Rat IgG1 kappa Isotype Control, eFluor 450

Catalog Number:

48-4301-80

In this experiment T cells were extracted from the spleens and lymph nodes of C57B/6 mice using single cell suspensions and negative selection columns, and were then subsequently cultured for 5 days in their respective cytokine milieu's in order to polarize them to a Th1, Th2, or Th17 phenotype. The Rat IgG1 kappa Isotype Control was used to differentiate non-specific background signal from specific antibody signal

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

C57B/6 CD4+ T cells

Primary Incubation

0.1ug/1.0x105 cells for 1 hour

Blocking Agent

20ug/ml gamma globulin

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

BD LSRII Flow Cytometer

Results Summary

The Rat IgG1 kappa Isotype Control was very effective in controling for background signal fluorescence.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

This control is very easy to use

The Bad

N/A

The Bottom Line

This is a great control to use at it is easy to optimize and is relatively inexpensive.

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