Immunofluorescence Imaging for Amplified CD11c

August 14, 2018

Molecular Biosciences
University of Texas Austin
Lab Technician

Overall

Quality of Results

Ease-of-Optimization

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Company:

Biolegend

Product Name:

CD11c N418 Antibody

Catalog Number:

117302

Our laboratory studies T Cell Acute Lymphoblastic Leukemia (T-ALL), a cancer of developing T cells. In order to examine the composition and organization of the tumor microenvironment, our lab utilizes a wide variety of markers to identify specific cellular subsets of the tumor microenvironment. We picked this antibody for immunofluorescence imaging knowing that it would characterize the architecture of the tumor microenvironment in T-ALL in our mouse model. We wanted to identify CD11c+ cells with IF imaging

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Lung, spleen, thymus

Primary Incubation

Overnight at 4 C

Blocking Agent

TNB blocking buffer, perkin elmer

Secondary Incubation

Perkins Elmer Biotin Amplification Kit, biotynil tyramide 1:200 for 5 min at room temperature, strepavidin HRP 1:200 for 30 min at room temperature

Tertiary Incubation

Streptavidin AF-588 (Life Technologies)

Detection

DMi8 Leica Microscope

Results Summary

This worked great for IF imaging! Be warned that you must amplify to get a good signal.

DOI or PMID #

Not yet published

Additional Notes

N/A

Related Categories

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Summary

The Good

Works great for me!

The Bad

I would recommend using an amplification kit for IF imaging. If you are in an organ with a lot of endogenous biotin (liver, lung, kidney, among others) then you need to block endogenous biotin. In the thymus or spleen, no endogenous biotin blocking is needed

The Bottom Line

Became a staple of my research, just make sure that you amplify it or you won't get much!

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