Anti CFTR Antibody For Fluorescence And Western Blotting

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Institute of Protein Biochemistry, Italian National Research Council, Napoli
Postdoctoral fellow

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Company:

R&D Systems

Product Name:

Human CFTR C-Terminus Antibody

Catalog Number:

MAB25031

Untagged CFTR is expressed in HeLa cells and was stained using the Human CFTR C-Terminus Antibody (MAB25031). The antibody stains CFTR specifically and it is good for immunofluorescence.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Hela cells overexressing CFTR

Primary Incubation

Primary antibody diluted 1:200 in 1% BSA- PBS and incubated for 1 hour at room temperature.

Blocking Agent

5% BSA prepared in PBS

Secondary Incubation

Coverslips were incubated for 1 hour at room temperature (in a dark chamber) with Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate (Thermofisher #A-11029) 1:400 diluted in 1% BSA- PBS.

Tertiary Incubation

None

Detection

Coverslips were inverted onto a glass slide with a drop of mounting media and Imaged using Zeiss LSM 700 confocal fluorescence microscope

Results Summary

Antibody stains the un tagged CFTR. The CFTR expression on the PM can be appreciated using this staining protocol.

Additional Notes

Fixed with 4% Para-formaldehyde and cell are permeabilized by inncubating the coverslips in 0.3% Triton X-100 in PBS at RT for 15 minutes

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Summary

The Good

Immunostains specifically CFTR

The Bad

Using in western blotting shows some nonspecific recognition of proteins.

The Bottom Line

Good reagent for immunofluorescence of CFTR

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