Anti-Psoriasin antibody [47C1068] (ab13680)

Hi there, I am planning to make immunoaffinity column with this anti-psoriasin antibody. Do you have any data about making Immunoaffinity column (antibody affinity chromatography) ? Which antigenic determinant (epitope) of psoriasin is recognized by this monoclonal antibody ? Epitope= the part of an antigen that is recognized by the antibody.

ANSWER:

Thank you for contacting Abcam. The epitope region of ab13680 has not been mapped and so I cannot inform you as to where the antibody actually binds. As for making an immunoaffinity column

1. Can you please detail this; are you observing differences between different batches or variability as the result of repeats from the same experiment using the same antibody batch? Can you tell me in what was was the result inconsistent. Was low signal observed or staining that was inconsistent?

I am observing differences between different batches. Using the same dilution, the intensity of my results are noticeably different from the first batch of antibody I received. I experienced low signal when using the same dilution of 1:100 between the different batches.

2. I would also appreciate it if you could provide me with details of the antigen retrieval approach that you are using; duration, approach etc.

The slides were boiled in target retrieval solution byDakocytomation. The solution which is 10x is diluted to 1x and boiled in a pressure cooker for 10mins at 120 celsius.

3. Have you been successful using this approach with other antibodies?

YES, WITH THE FIRST BATCH OF ANTIBODY, MY RESULTS WERE GOOD.

4. Are the samples that you are using human?

HUMAN SAMPLES

ANSWER:

Thank you for getting back in touch with and for providing me with further details.

The difficulties that you have been having are clearly lot related problems. I would like to offer you a free of charge replacement vial of a different lot. However, unfortunately the lot number that you received on order 152018 (PO number: 0001862788) was the same as the lot currently in stock. I would therefore like to offer you a credit note to the value of one vial of this antibody. If this is acceptable please email me and I will request that our accounts department processes your order details.

I look forward to hearing from you.

BATCH NUMBER 47C1068 ORDER NUMBER 00018627

DESCRIPTION OF THE PROBLEM low signal. not consistent result as first order of the antibody.

SAMPLE tumor tissues

PRIMARY ANTIBODY 1:100 dilution. tried 1:50 dilution but signal still not very strong. ran a pos. control with the rest of the slides.

DETECTION METHOD fast red tablet in substrate buffer.

POSITIVE AND NEGATIVE CONTROLS USED positive controls used

ANTIBODY STORAGE CONDITIONS -20 celsius

FIXATION OF SAMPLE parafin embeded tissues on slides

BLOCKING CONDITIONS target solution from [another company]

SECONDARY ANTIBODY [another company] kit using polymer labelled AP as secondary antibody.

HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? No

WHAT STEPS HAVE YOU ALTERED? no steps altered. just ordered a new antibody

ANSWER:

Thank you for your enquiry.

I am sorry to hear that you have had variable results using this antibody. I have read your questionaire and I have a few comments.

I am most interested in the variability that you have been observing.

1. Can you please detail this; are you observing differences between different batches or variability as the result of repeats from the same experiment using the same antibody batch? Can you tell me in what was was the result inconsistent. Was low signal observed or staining that was inconsistent?

2. I would also appreciate it if you could provide me with details of the antigen retrieval approach that you are using; duration, approach etc.

3. Have you been successful using this approach with other antibodies?

4. Are the samples that you are using human?

I appreciate your patience and look forward to hearing from you.