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Dear Sr./ Mrs.,
I am writing to you to discuss a problem we have just had with two
antibodies purchased at your company, Rictor (AB104838) and Raptor
We have just performed immunofluorescence staining on mouse cerebral
tissue The staining did not work out and was similar to a negative
control (without primary antibody and plus secondary antibody).
The protocol that we used was the following:
Sections were washed twice with PBS and incubated with NH4Cl 50 mM
during 30 min. Tissue was permeated along 20 min by treatment with PBS
containing 0.5% Triton X-100 and blocked during 1 h at room
temperature in PBS plus 0.2% bovine serum albumin, 0.2% glycine, 0.2%
sodium azide, 0.5% Triton X-100 and 5% normal goat serum. Slices were
incubated overnight at 4°C with the corresponding primary antibodies
in a buffer containing PBS plus 0.3% Triton X-100, 0.2% bovine serum
albumin and 0.2% sodium azide. After primary antibody incubation,
slices were washed in PBS twice for 10 min and incubated 2 h at room
temperature with subtype-specific fluorescent secondary antibodies.
After two consecutive 10 min-washes, slices were mounted with Mowiol
on silane-coated slides.
Primary antibodies were diluted 1:200 in the case of Rictor, and and
1:100 in the case of Raptor, as recommended on the data sheet.
We are not sure if we should follow a specific protocol to get the
staining work out fine.
Thank you very much,
Asked on Mar 26 2012
Thank you for contacting us.
I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
In order to better understand the problem, I would like to send you a questionnaire with some additional questions about the protocol used, as well as the order details. All this information is crucial to help us understand the possible causes of the problem. Any images are highly appreciated.
Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. If no suggestions can be made to the protocol, and the antibody was purchased within the last 6 months, it is covered by our Abpromise guarantee, and therefore I can offer you a free of charge replacement or a credit note for it.
I look forward to receiving your reply. Please do not hesitate to contact us if you need any more advice or information.
Answered on Mar 26 2012