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Anti-DDAH1 antibody (ab2231)

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1 question for ab2231

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Question 1

Friday 20-June-2003

Hi! So, we had a customer call today who is using ab 2231 in both human and rat aortic homogenate. He has loaded from 100 ugs to 1 mg of protein per lane diluted the antibody 5 ugs/ml and incubated overnight. He sees a band at 32kDa but nothing at 40 kDa. It says on the datasheet that the predicted MW is 33 kDa but runs at 40 kDa do you have the peptide available for them to run a blocking study? The other issue is that they see this band in their negative control lane (with BSA). Otherwise, this looks to be an issue with the antibody.

ANSWER:

  

According to the datasheet, this antibody is thought to run at 40kDa due glycosylation. It could be that the protein is running at 32kDa in their experiments as the protein in their system is not glycosylated. However, I am concerned that their negative control is also showing a band at the same size. We do not have the peptide here for them to try but I will ask the originator of the antibody if they have the blocking peptide available. It would help if the researcher could contact us directly and provide a blot image. Are they just loading BSA in their negative control lane? Please ask them to contact us by e-mailing technical@abcam.com and attaching a blot image to the e-mail.

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