Anti-SOCS1 antibody (ab3691)

I just would like to know whether you are still dealing with the problem I mailed you. [CCE1113967]

ANSWER:

I apologize for the delay in finding out the results of the test of the antibody, I am pleased to inform you that we have re-tested this lot against our new lot and found that its activity has decreased over time. We would therefore like to offer you a replacement vial of the new lot, which was just tested and gave a good band at 37kDa in human spleen lysate. I have added the image of the test on the datasheet immediately for you to see.

I am very sorry it has taken unusually long to resolve this matter. I would appreciate if you could confirm if you would be interested in trying the new lot, or if you would prefer a refund on your order 250110.

Thank you very much in advance,

I would like details on how the HEK cells were treated with GH in order to get that 24 kDa band.

ANSWER:

Thank you for your enquiry. The HEK cells were incubated with 500 ng/mL human growth hormone for 6-12 hours.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

thanks I have a question concerning the ab3691 Abody (Socs1). I should get a band with Hek293 around 24KDa in presence of GH. I guess GH stand for Grow hormone. does the 293 has to be GH treated to get the 24KDa band. How so you treat the 293 cell with GH?

thanks

ANSWER:

Thank you for your enquiry. The stimulation conditions for the HEK293 cells seen in the Western blot image are 500 ng/ml human growth hormone.

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Thank you for your immediate response. My next question is whether you are aware of any available proteins that could work as positive controls for ab3691 (SOCS1 antibody) and ab14939 (SOCS3 antibody) for quantitative results in an ELISA assay in human samples (liver, muscle, adipose tissue)?

ANSWER:

Thank you for getting back in touch with me.

As mentioned in my previous email I consider SOCS1 peptide (199-211) (ab23167) suitable as a positive control standard for ab3691 as this is the very peptide sequence that the antibody was raised against.

With regards mouse monoclonal [SO1] to SOCS3 (ab14939) we do not have a positive control in our catalogue. However, following a brief "Google" search I can tell you I found an epitope tagged recombinant human form of SOCS3 available at the following link;

http://www.axxora.com/socs-related_products-ALX-201-334/opfa.1.1.ALX-201-334.1872.4.1.html

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Thank you very much for your reply. I need to use for SOCS 1 (Ab 3691) a SOCS 2 (Ab 3692) some positive controls, such as whole-cell lysates (WCEs), which are used directly as positive controls employing Western blots. Please could you give me some advice?

Thank you.

ANSWER:

Thank you for your enquiry.

Unfortunately we do no have a recommended positive control for SOCS1 antibody (ab3691). However, the customer that left a favourable Abreview used a human cell lysate - whole cell (human malignant melanoma cells). Furthermore Ramos or RAW cells are recommended according to the literature.

With respect to SOCS2 antibody (ab3692) we recommend the use of lysates of GH treated HEK 293 cells transfected with GHR. Furthermore in the following publication that has used this antibody C57BL/6 mice overexpressing a SOCS2Tg were employed for use by IHC.

Scott HJ et al. Differential effects of SOCS2 on neuronal differentiation and morphology. Brain Res 1067:138-45 (2006). PubMed: 16360125

I hope this information helps, please do not hesitate to contact us if you need any more advice or information.