Anti-CTLA4 antibody (ab9984)

The datasheet for this antibody was incorrect, the immunogen and species reactivity were listed as human, but actually were mouse.

ANSWER:

I apologize for the confusion with this antibody. Unfortunately I do not know where the mistake on the datasheet came from, but I have already updated the datasheet to be sure this problem does not happen again.

As agreed over the phone, I am sending you a free of charge vial of ab19792 as a replacement for one of your orders, and refunding the other two orders. The order number for the free of charge vial is 137206. You should get a separate email with shipping details for this.

For any questions regarding the refunds, please contact our accounting department at accounts@abcam.com.

I sincerely apologize for this problem. I am sorry that this happened. I hope that ab19792 works for in your Western blot, and encourage you to submit an Abreview of both antibodies (we accept negative reviews as well as positive).

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 027T0022

DESCRIPTION OF THE PROBLEM No bands for CTLA-4 Too many bands for Smurf2

SAMPLE Human melanoma cell extract - RIPA

PRIMARY ANTIBODY 1:1000 in Blocking buffer at 4 C overnight 1:500 in Blocking buffer at 4 C overnight Wash 3 times 5 min

DETECTION METHOD ECL

POSITIVE AND NEGATIVE CONTROLS USED Don't have; looking for published bands from Ab from other companies

ANTIBODY STORAGE CONDITIONS 20ul aliquots at -20 C

SAMPLE PREPARATION RIPA + Complete protease inhibitors

AMOUNT OF PROTEIN LOADED 20 ug

ELECTROPHORESIS/GEL CONDITIONS 8% SDS-PAGE

TRANSFER AND BLOCKING CONDITIONS Transfer: Tris Acetate SDS with 20% Methanol Blocking: 5% Nonfat dried milk in TBS-T

SECONDARY ANTIBODY 1:5000 in blocking buffer 2hrs at room temp 1:2000 in blocking buffer 2hrs at room temp Wash 1x 15min and 3x 5min each

HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

WHAT STEPS HAVE YOU ALTERED? For CTLA-4: more protein primary antibody conc. and incubation time secondary Ab conc and time

For Smurf2: more washing steps longer blocking time less primary antibody

ADDITIONAL NOTES These antibodies were purchased more than 90 days ago, but considering how many antibodies we order from abcam, I hope that you can refund the money to us or credit our account. We will surely order many more antibodies from abcam in the future.

Thank you very much for your help.

ANSWER:

Thank you for the details that you have provided. At this point I would suggest using positive controls with these antibodies. For ab17919, HepG2 lysate is recommended and for ab9984, we would suggest using a type of human lymphoid tissue as the highest levels of CTLA4 are found in lymphoid tissues.

I hope this helps. Please contact us again if you have any additional questions.

I was wondering if this antibody can be used for immunoprecipitaation experiments

ANSWER:

Thank you for your enquiry. To our knowledge, this antibody has yet to be tested in IP. All tested applications are specified on Abcam product datasheets. If you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher. If you have any more questions please contact us again.

Can this antibody detect the extracellular domain of CTLA4. This is because the protein product I am attempting to analyse contains only the extracellular domain.

SAMPLE CHO producing a fusion protein product containing the extracellular domain of CTLA4

TYPE OF ELISA Sandwich

ANSWER:

Thank you for your enquiry. Based on the sequences, the antibody should recognize the extracellular domain of CTLA4. The antibody was produced from an E. coli derived protein, so we do not know for sure if the detection may be decreased by the fusion protein produced in CHO cells.

Generally, we would expect the antibody to recognize the target protein in your sample, but we are not certain of the lower limits of detection.

This antibody is reported for elisa. Can you give me some more details about this application...which antigen has been used....which secondary antibody...regards.

ANSWER:

Dear Dr. D'Erasmo,

All the information we have regarding this antibody is located on the online datasheet. The supplier of ab9984 was not able to provide any additional details that we do not already have.