Anti-Cytochrome P450 Reductase antibody (ab13513)

Thank you for bringing this to our attention and for sending your protocol and IHC images. I am sorry to see that the antibody is giving you trouble. It is unlikely, but possible, that the wrong antibody was delivered to you. We have not received other complaints about the antibody so I do not think there was a labeling mix-up. The antibody may itself be defective, at least with these mouse samples. I am wondering if the glutaraldehyde that you include in the fixative might be creating some of the background but your negative control appears relatively clean. Some people will add glycine at 0.3M to the blocking solution to quench unreacted aldehydes. In any case, there is no staining of the ER, as far as I can tell. Can you tell me how long your samples were fixed, how much antibody you applied (concentration or dilution), and if you are confident that the secondary antibody is effective? For a replacement or refund or credit, I will also need the order reference number, either Abcam's or the PO number. I look forward to your reply and to resolving this issue.

Thank you for contacting us with your question. Ab13513 is provided as whole antiserum and is not a purified antibody. Some of our other cytochrome P450 reductase antibodies are affinity purified (such as ab39995) and we also have purification kits that can be used to isolate the IgG molecules from the antiserum- https://www.abcam.com/Antibody-Serum-Purification-kit-1-purification-ab109208.html I hope this information will be useful in determining whether this antibody is appropriate for your studies. If you have any further questions or need any other information please let me know.

Thank you for contacting us. For ab23677, LTA4 hydrolase, I suggest either a neutrophil or monocyte lysate. We carry THP1 lysate, ab7913, which is a human monocytic leukemia. For a negative control, LTA4 was detected in epidermal keratinocytes and endothelial cells but not whole skin in the following paper: Leukotriene A4 hydrolase in human skin. J Invest Dermatol. 1994 Feb;102(2):253-7. PMID: 8106755 Our lysate ab30166 may be suitable as a negative control but I cannot guarantee you will not see some reactivity. For the cytochrome P450 enzymes, I suggest liver lysates. A positive control for ab4236 is human liver: ab29889. For ab13513 and ab3573, rat liver lysate ab29413 will be appropriate. I am reluctant to recommend a negative control since information regarding absence of expression is difficult to come by but since CP450 is associated with the ER, cytoplasmic fractions of liver lysates (which do not contain membrane) may be useful. For human liver, this would be ab29875, and for rat liver ab29409. You should also have a look at the references listed on the datasheets to see how other researchers have used the antibodies, though the references for the LTA4 are general and not specific to the antibody. I hope this helps. Please do not hesitate to contact us if you have any other questions.

Thank you for your enquiry. I was able to obtain a Western blot image, which I have added to our datasheet. The antibody was used in WB in this paper, http://cancerres.aacrjournals.org/cgi/content/full/60/13/3550 I hope this information helps. Please do not hesitate to contact us if you need anything further.