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As discussed over the phone earlier today, we have encountered a problem with the Abcam pERK Ab.
Our initial working up of this was carried out using Ms mAb to ERK1 +ERK2, [MAPK-YT] (phospho T185), 50ul (5.2mg/ml), AB50011, Lot: GR121-7.
The results of this were good, reasonably clean, nuclear staining on our colorectal cancer TMA sections and control kidney sections.
We determined the optimal dilution to be 1:50.
The TMA image can be viewed at http://126.96.36.199/Research_4/Clinical_Trials/Susan_Richman/Gaya/190712/view.apml?
The file is 183507.svs and can be opened with Webscope or Imagescope if you download the free software.
I should be able to send you an image of the slides stained with lot number GR121-8 tomorrow or Monday as a comparison.
AS we had insufficient Ab left, I re-ordered it, and was sent a vial identical to the above, with the exception of the lot number being GR121-8, instead of -7.
The resultant staining with this new lot showed absolutely no nuclear staining whatsoever. There was minimal membranous staining at best. The protocols used were identical in every way, except the Ab.
This is a real concern as there is obviously lot to lot variations and we have unfortunately wasted valuable clinical trial material. Have such problems been encountered elsewhere?
We are limited by time on this project, so hope to resolve this ASAP. Would you be able to look into sourcing more of the lot GR121-7 for us, which we could re-test on colorectal control material before sacrificing valuable clinical material again.
Look forward to hearing from you in due course,
Asked on Jul 27 2012
Thank you for taking the time to contact us. You enquiry has been forwarded to the scientific support team. I am sorry to hear you have had difficulty obtaining satisfactory results from this vial of antibody.
I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.
In answer to your question regarding the lot numbers, we do have lot GR121-7 still in stock. However, reviewing the details I can confirm that our lots GR121-7 and GR121-8 are the same original batch from the producer, the vials have just come to us in different deliveries. I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch.
I would also like to reassure you that this antibody is tested and covered by our 6 monthguarantee for IHC-P (paraffin embedded) and human samples. Before deciding how to proceed, I would like to investigate this particular case further for you, and also obtain some further information for our quality records.
In order to do this, I have enclosed a questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily. I would appreciate if you are able to provide some images which will help us to assess the results. It would be helpful if these could be attached to the reply email in PDF or JPEG format.
In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.
Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.
Purchase order number
or preferably Abcam order number:
Antibody storage conditions (temperature/reconstitution etc)
Description of the problem (high background, low signal, non-specific satining etc.)
Sample (Species/Tissue/Cell Type/Cell Line etc.)
Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.)
Antigen retrieval (Enzymatic method, Heat mediated technique etc.)
Blocking conditions (Buffer/time period, Blocking agent etc.)
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Positive and negative controls used (please specify)
Optimization attempts (problem solving)
How many times have you tried the IHC?
Have you run a "No Primary" control?
Do you obtain the same results every time?
What steps have you altered?
We would appreciate if you are also able to provide and image which would help us to assess the results
Answered on Jul 27 2012