Product Review: PYCR1 antibody – middle region (ARP57751_P050) using whole extracts from Human fibroblasts and 293T cells using Western Blot

Product Page Link: PYCR1 antibody – middle region (ARP57751_P050)

Data provided by Dr. Samuel S.W. Szeto of Human Embryology and Genetics Lab, Institute of Medical Biology, A*Star, Singapore

How do Aviva’s reagents play a role in your experimental goals?

Western blots are a critical component of the research we perform as we are trying to determine the molecular mechanisms of various genetic disorders. We need to be able to assess the status of our target proteins.

How many different experimental trials were conducted using the antibody sample? One trial western was done with each of the four antibodies provided.

What type of experimental sample are you using and how did you prepare it?  Samples were whole cell extracts from patient fibroblasts (mutant and WT for PYCR1) and from 293T cells transfected with either PYCR1, PYCR2 or PYCR-flag. For the samples from patient fibroblasts, samples were TCA precipitated because they were quite dilute and I wanted to load more protein into the gel. 10 ug for controls and 100 ug for patient fibroblast samples.

What applications did you test the antibody in? Please include dilutions of the primary and secondary reagents. Western blots only. For primary, I used at the dilution of 1 ug/mL. I was only given enough material to use at this dilution. For our secondary we used Donkey anti-rabbit HRP at 1:3000.

How did you store the antibody after re-suspension? In 5% milk, 1xPBS + 0.1% NP-40. Frozen at -20 deg.

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

I followed the immunoblot protocol for cell lysates on your webpage. I used SuperSignal West Pico, Dura and Femto CL reagents from pierce to do the detection.

 Would you use this antibody in future experiments? I am considering using the Aviva antibodies.

Do you believe the information about the reagent on Aviva’s website is correct?

The information provided is correct.

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