Anti-MRP2 antibody [M2 III-6] (ab3373)

I can confirm that anti MRP2 antibody ab3373 does not cross react with other MRPs.

Thank you for your reply and for kindly providing the requested details.

I look forward to hearing from you with the results of the next experiment. i can recommend it would be beneficial to consider a reducing gel. This will ensure the protein is in the correct conformation for detection by the antibody. Although the other antibodies work well using a non reducing gel, individual antibodies detect individual epitopes and so will sometimes require individual optimization.

I hope this will be helpful. Please do not hesitate to let me know how you get on with the reducing gel. I look forward to hearing from you.

Thank you for taking the time to complete our questionnaire. The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimise the results from these antibodies. I would also appreciate if you can confirm some further details:

1. Please confirm the order numbers and dates of purchase.

2. Could you confirm which lysis buffer was used? I can recommend to try RIPA buffer if not already tried. This should provide a suitable protein preparation.

3. We recommend samples should be reduced and denatured and run on a reducing gel in WB unless stated otherwise on the datasheet. This will ensure the protein is in the correct conformation to run at the correct molecular weight and be detected by the antibody.

4. I can recommend to consider trying BSA rather than milk to block. Changing blocking agent can sometimes help to improve results.

5. Could you confirm if the transfer to the membrane and quality of the samples been assessed using a loading control?

6. Are the current vials of secondary antibody and detection kit working well with other primary antibodies?

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from these antibodies.

I am sorry to confirm that because we carry over 90,000 products, it is regrettably not feasible for us to stock small sample sizes of our products to send to customers. If you are concerned about the secondary antibody, I can recommend to consider contacting the supplier of the secondary. I notice that MRP4 antibody [M4I-10] (ab15602) is a rat IgG2a, do you know that the secondary antibody will detect this isotype?

ab3373 Anti-MRP2 antibody [M2 III-6] should be working in WB, it has been tested successfully. Particularly if it has worked in the IHC experiments which shows the antibody is active, it is unusual this is not working in WB. Again, this is a mouse IgG2a isotype. Will the secondary antibody used in WB detect this, was this the same secondary used in IHC-P?

I would like to reassure you that all these antibodies are tested and covered by our 6 month guarantee for WB and the species that are listed on the individual datasheets. 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.

I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

Thank you for your time and cooperation. I look forward to receiving the completed questionnaire.


Order Details
Antibody code:
Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background
Lot number
Purchase order number
or preferably Abcam order number:
General Information
Antibody storage conditions (temperature/reconstitution etc)
Description of the problem (high background, wrong band size, more bands, no band etc.)
Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
Amount of protein loaded
Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
Transfer and 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)
Detection method (ECL, ECLPlus etc.)
Positive and negative controls used (please specify)
Optimization attempts (problem solving)
How many times have you tried the Western?
Have you run a "No Primary" control?
Yes No
Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?
What steps have you altered?
Additional Notes:
Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to assess the results.