E1A Antibody Good For Immunocytochemistry

November 14, 2016

Center for Hearing & Deafness
University at Buffalo
Principal Investigator

Overall

Quality of Results

Ease-of-Optimization

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Company:

Abcam

Product Name:

Anti-Adenovirus E1A antibody [0.N.7]

Catalog Number:

ab33183

We use the 12S E1A gene of adenovirus expressed in a replication-incompetent retrovirus as a means to generate immortalized cell lines from various rat and mouse tissues. One way to test whether our cells of interest have been infected with the virus is to stain for 12S E1A in cell lines that have been exposed to the retrovirus. We chose this particular anti-E1A antibody because it is listed as being useful for multiple applications, including IHC and western blot. Here, we demonstrate the usefulness of the antibody for immunofluorescence.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Cytospin of the Psi2 packaging cell line that produces the 12S E1A expressing retrovirus

Primary Incubation

One hour at room temperature at 2 ug/ml in PBS-Tween

Blocking Agent

None

Secondary Incubation

Vector Dylight 549 anti-mouse antibody (D12549) used at 5.0 ug/ml at room temperature for 45 minutes

Tertiary Incubation

Vectashield mounting medium containing DAPI counterstain

Detection

Fluorescence microscopy- Nikon microscope with SPOT camera

Results Summary

The Psi2 packaging cell line was expected to have a great deal of immunofluorescence due to the production of retrovirus expressing 12S E1A. This was evident in the attached figure, with red immunofluorescence corresponding to E1A immunoreactivity. DAPI is seen in blue as a counterstain for cell nuclei.

DOI/PMID #

N/A

Additional Notes

None

Related Categories

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Summary

The Good

Very easy to optimize, good detection, low background

The Bad

None noted.

The Bottom Line

In summary, we recommend this 12S E1A antibody from Abcam. It is a mouse monoclonal that can be used in combination with other primary antibodies for double labeling studies.

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