Anti-CISH antibody (ab5923)

The customer would like to know if the antibody was tested in IP on tissue lysate or with recombinant protein. Do we have more information and an image illustrating the testing.

ANSWER:

The antibody has been tested in IP against endogenous protein, not recombinant protein. It was raised against a synthetic peptide corresponding to 170-185 amino acids of human/mouse/rat CIS-1 therefore if your recombinant protein of interest covers this area, it should be applicable.

Customer is using this antibody in IP and would like to have the exact protocol that was used when this antibody was tested in IP? Also, what cell line was used for testing?

ANSWER:

Thank you for your enquiry and patience. I have just heard back from the originator of this antibody regarding your question. Ab5923 was tested in IP using mouse embryonic cells. The IP protocol that they followed is below:

- 2 ul cell lysate -add primary antibody -incubate o/n at 4C -add Protein A agarose beads (20 ul) -incubate 1-3 hr at 4C -micro-centrifuge for 30 min at 4C -wash pellet 2x, discard supernatant -wash pellet again 2x -add 200 ul PBS -micro-centrifuge pellet and discard supernatant -add 50-100 ul SDS -boil 5 min -micro-centrifuge 1 min -save supernatant -load sample (15-30 ul) onto 10% SDS-polyacrylamide gel. -perform Western blot

Customer purchased ab5923, however it is not a SOCS1 ab but a CIS1 ab.

ANSWER:

As I explained to you on the phone, there was a mistake on the datasheet for ab5923 - it is a CIS1 antibody, not a SOCS1 antibody. I apologize for this mistake and the datasheet has been updated to reflect this information.

I can of course give you a refund or send you another antibody free of charge. Ab9870 is a SOCS1 antibody that may be of interest to you. Please let me know which option you would prefer.

It states that ab5923 detects a band of 47kD in WB when the predicted MW of SOCS1 is 24 kDa. Ab3691, also a SOCS 1 antibody, detects a band at 24 kDa - why?

I still have not heard a response to the enquiry reference CRM372981 and it has been a week. This has slowed down my research and questioned everything completed thus far because it is not certain whether the antibody (ab5923) actually recognizes SOCS-1. I would like to try another SOCS-1 antibody , however, I am hesitant because I am not pleased with ab5923 and I would like to know that the antibody I am interested in, ab9870, definitely recognizes SOCS-1. Please let me know if you have any information.

ANSWER:

Thank you for your patience. As ab5923 was originated outside Abcam I had to contact the originator regarding your question and have just heard back.

There was a mistake on the datasheet for ab5923 - it is a CIS1 antibody, not a SOCS1 antibody. I apologize for this mistake and the datasheet will be updated to reflect this information.

I can of course offer you a refund or an alternative antibody free of charge. Ab9870 is a SOCS1 antibody, as well as ab3691. Please let me know what you would like.

Hello, I'm interested in your SOCS1 anti-bodies : ab3691 and ab5923 : I want to know which antibody you used for capture and which one for detection in ELISA applications. Thanks for your response and best regards.

ANSWER:

Thank you for your enquiry and your interest in our products.

We would like to confirm that these two antibodies (ab3691 and ab5923) have been characterized for ELISA application. However, they have not been tested together in a sandwich ELISA system.