Anti-COX2 / Cyclooxygenase 2 antibody (ab6665)

Has this antibody been tested in IHC? Are there any references?

ANSWER:

Thank you for your enquiry. To our knowledge, this antibody has yet to be tested in this application. 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.

The only references were are currently aware of are the following (the Pubmed links are on the online datasheet):

Tong M & Tai HH Induction of NAD(+)-linked 15-hydroxyprostaglandin dehydrogenase expression by androgens in human prostate cancer cells. Biochem Biophys Res Commun 276:77-81 (2000). PubMed: 11006085

Guan Z et al. Interleukin-1beta-induced cyclooxygenase-2 expression requires activation of both c-Jun NH2-terminal kinase and p38 MAPK signal pathways in rat renal mesangial cells. J Biol Chem 273:28670-6 (1998). PubMed: 9786861

I have a question regarding two antibodies I have ordered from recently:

1) By using the antibody ab6276 (beta-actin) in Western Blotting I see a band at around 80 kDa. Is beta-actin dimerazing or what could this band be?

2) Using the ab6665 (COX-2) antibody I see a lot of additional bands, some are even bigger that the COX-2 band at 70 kDa. There are bands at 32, 40 (very big), 53, and 60 kDa. Why is that and what can I do against it?

I would appreciate your help a lot.

ANSWER:

Thank you for your patience. I was away on vacation and thus could not get back to you earlier.

1) Using ab6276, you are most likely detecting a dimer. I would suggest trying the antibody with different samples to ensure that the antibody detects the 42 kDa band.

2) It seems that you are detecting some non-specific bands in addition to the Cox2 band at 70 kDa. To try to get rid of these bands I would suggest decreasing the amount of primary antibody used and also decrease the primary incubation period. Also, change the concentration of the secondary antibody and make sure to do a secondary antibody control (no primary).

Customer is using ab6665 on the following cell line: SKBR3 (breast cancer cell line) and is seeing a very strong band at 50 kDa, a fainter band at 70 kDa and some other faint bands as well.

ANSWER:

The originator of ab6665 informed me that the band close to 70kDa should be the specific band. The others are non-specific. This result is very consistent with what they have previously seen. They don't have a recommendation for a positive control but suggested searching for a cell lysate proven to have cMyc tagged protein.

Thanks for your help with my questions regarding COX-2, but I have one more! Since the lysates that were used as positive controls for this antibody aren't available for purchase, I was wondering if you would be able to recommend me a lysate that you do carry that I may be able to use. I checked one of the references listed for this ab, but it did not mention your company specifically. I wasn't sure if perhaps these researchers (Min Tong and Hsin-Hsiung Tai) licensed it to Abcam for commercial production. If you would be able to recommend a positive control for this ab, that would be great. I very much appreciate any assistance you're able to provide.

ANSWER:

As you probably know Cox-2 (Cyclooxygenase-2) is an inducible enzyme that is normally absent from cells, however, in response to growth factors, tumour promoters and some cytokines, it undergoes a rapid and transient expression.

We are very sorry to inform you that we do not have positive controls for this particular antibody.

Dear Abcam, I want to do IHC of COX1 & COX2 in Bat uterine tissues. Could toy please help me in this matter.Do you know anybody who can sevd me the protocol and kit for my assay. I dont have too much of fund, so I need to be very careful for spending also. Thanks and regards.

Dr Pranab Lal Pakrasi

ANSWER:

I don't recall that any of our products have been tested for cross reactivity with bat! ab6665 has not yet been tested in IHC. I suggest that you take a look at the IHC and antigen retrieval protocols on our the Abcam website. This is the only antibody against COX-2 in our catalog, and currently, we do not stock antibodies to COX-1.