Product Review: TNKS antibody – middle region (ARP33755_P050) in mouse MEF lysate using WB

Product Page for TNKS antibody – middle region (ARP33755_P050)

Researcher: Daolin Tang, University of Pittsburgh Medical Center
Application: Western Blotting
Species+tissue/cell type:
Lane 1-6: 30ug mouse MEF lysate
Primary antibody dilution: 1:200
Secondary antibody: Anti-Rabbit-HRP
Secondary antibody dilution: 1:2000

 

 

 Questionnaire:
 How would you rate this antibody on a scale from 1-5 (5=best) nad why? 4.5
 Would you use this antibody in future experiment?  Yes. I have ordered the other vial from your company
 Have you used another antibody which has worked in your application?  No
 Do you believe the information about the reagent on Aviva’s website is correct?  Yes. But there do have some problem with the antibody information, such as TERF1 antibody (OAAB01445), the mouse protein size is ~46 kd, but the size on the data sheet ( image 2) is around 72kd .
 If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?  Yes. I have ordered the other new vial from your company
 How did you store the antibody after re-suspension?  4 degree.
 Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):  Mouse, MEFs, 30ug
 How many different experimental trials were conducted using the antibody sample? 4 to 5
 How was this sample prepared?  I used the Tris/ SDS lysis buffer.
 Primary antibody dilution and incubation time:  1:200 4 degree overnight
 Secondary antibody used and dilution and incubation time:  Anti rabbit from cell signaling 1:2000
 What controls were used in your experiment (positive/negative)?  I didn’t use positive control because the signal is very specific and close the predicated size of the protein.
 Please include your detailed WB Procedure/Protocol here:  Mouse MEF cells
Blocking buffer: 5% milk
1st antibody: 1:200 at 4 degree overnight
2nd antibody : 1:2000 at RT 1hr
Washing buffer: PBST
Test with ECL from Pierce
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