Customer Reviews for ASTN2 Antibody (ARP46963_P050) tested with mouse cerebellar granule cells in Immunofluorescence

CAT# ARP46963

submitted by:
Hourinaz Behesti
Rockefeller University

How many different experimental trials were conducted using the
antibody sample?

2 immunos, 1 WB

What type of experimental sample are you using and how did you
prepare it?

Human fibroblasts, mouse cerebellar granule cells

What applications did you test the antibody in? Please include
dilutions of the primary and secondary reagents.

WB: did not appear to work
ICC: 1:100 dilution. Alexa 495-conjugated anti Rabbit used as secondary

What controls were used in your experiment? Please include your
positive control.

Positive control: MC7 cells that express Astn2, negative control: no antibody

For IHC, what antigen retrieval method did you use?

none, used 0.05% Triton in the block, and primary incubations

How did you store the antibody after re-suspension?

-20C

Please provide the protocol for your application procedure. Please
be as detailed as possible.

1-Block: 5% normal goat serum, 0.05% Triton in PBS 2-Primary incubation: 1:100 for 1 hr at RT
3-2×5 min PBS washes
4-Secondary incubation: 1:300 for 1 hr at RT
5-3×5 min PBS washes
6-mounted with antifade medium

Would you use this antibody in future experiments?

Yes

Please explain any problems you had with the antibody and/or
experimental results that Aviva can address in the future.

It does not detect the band at ~115 kDa for Astn2, which I am able to detect with our own home-made antibody.

If the antibody works, do you plan to use it in future experiments
or to publish your data? Why or why not?

Yes if I can verify that it binds to Astn2 specifically

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