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I am not able to get this antibody to work. I used mouse E12.5 and adult mouse uterine tissue. Both these tissues have confirmed Hoxa11 expression. I used wt, het and mutant tissue to discern the specific Hoxa11 protein band as it is not expressed in our mutant mouse. I used an anti-GFP antibody as a positive control. The GFP is expressed in mutant and het tissue. The control worked.Here is the protocol that I followed. Block: 1hr in 5%milk in TBST. Antibody incubation: 1:500 dilution in 5%milk TBST O/N at 4 degrees Wash 6 x 5min in TBST Secondary: Goat anti Rabbit HRP Wash 6 x 5min in TBST Chemiluminescence detection Attached you will find supporting documentation.
Asked on Aug 14 2012
Thank you for contacting Abcam regarding ab72591.
I am sorry that you have been experiencing difficulties with this antibody in WB. I have reviewed your protocol and image and I agree that this product is not working as stated on the datasheet. I am happy to offer a replacement or credit. Please let me know which you would prefer. Would you also include your order number with your reply?
I look forward to your reply so that I may assist you further. Please do not hesitate to contact us if you have any additional questions.
Answered on Aug 14 2012