Product Review: COL1A1 Antibody (AAS99599C) in Mouse blastocyte using IHC

Product Page for COL1A1 antibody-C-terminal region (AAS99599C)

Researcher: Ivan Bedzhov, PhD, Gurdon Institute, University of Cambridge
Application: IHC
Species+tissue/cell type:Species+tissue/cell type: Mouse blastocyte
Primary antibody dilution: 1:200
Secondary antibody: Anti-rabbit-Alexa 488
Secondary antibody dilution:1:200


Questionnaire: How do Antibody Verify’s reagents play a role in your experimental goals? How would you rate this antibody on a scale from 1-5 (5=best) and why? 5, clear and shARP signal. Would you use this antibody in future experiments? Yes Have you used another antibody which has worked in your application? Yes Do you believe the information about the reagent on Antibody Verify‚Äôs website is correct? Yes If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not? Yes, I plan to use it and evantually publish. How did you store the antibody after re-suspension? 4 degress Please explain fixation solution/method used (formalin, periodate-lysine-paraformaldehyde, acetone, etc.)? PFA How many different experimental trials were conducted using the antibody sample? 3 Primary antibody dilution, incubation time and temperature: At 1:200 Secondary antibody used, dilution, incubation time and temperature: Alexa488 From your IHC/ICC images, briefly explain the colors of each stain and counterstain: Green: Collagen, Blue: DAPI Did you use an antigen retrieval method? If so, please explain? No What controls were used in your experiment? Please include your positive control: Blastocyst outgrowth stained for Laminin Experimental Procedure/Protocols: Fix in 2% PFA in PBS.
Wash 3X PBS
Permeabilize 0.5% Triton-X-100 in PBS, 15 min
Wash 2X PBS
Primary antibody 1:200 in PBS, overnight, 4 degrees
Wash 3X PBS
Secondary antibody 1:200 in PBS, overnight, 4 degress or 2h RT
Wash 3X PBS

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