Immunofluorescence Using Transferrin Receptor

Cell & Tissue Engineering, Imec
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Invitrogen

Product Name:

Transferrin Receptor Monoclonal Antibody (H68.4)

Catalog Number:

13-6800

We conducted transferrin receptor (TfR) staining in primary human brain endothelial cells (HBMECs).

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Primary human brain microvascular endothelial cells

Primary Incubation

Dilution 1:100

Blocking Agent

Donkey serum

Secondary Incubation

Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 647, #A-31571 (dilution 1:500)

Tertiary Incubation

none

Detection

Confocal

Results Summary

Cells were fixed in 4% PFA, and permeabilized with 0,3% Triton x-100. The optimization takes some time and it's possible to observe high background signal, compromising the visualization of the signal. Cells were countedstained with Hoescht and Ve-Cadherin for detection of nuclei and tight junctions, respectively.

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Summary

The Good

Some signal can be detected

The Bad

High background signal

The Bottom Line

This antibody performs very well in WB; however, IF applications may require some time and optimization to achieve optimal results.

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