Anti-Oligodendrocyte Specific Protein antibody - Oligodendrocyte Marker (ab7474)

I bought the Anti-Oligodendrocyte Specific Protein antibody - Oligodendrocyte Markeron *******. We used for IHC-Fr staining and doesn’t work properly. The ****** I would like to know if we can have a refund or credit back for this antibody. Thanks. Have a nice day!

ANSWER:

Thank you for contacting Abcam.

Under our Abpromise we guarantee our antibodies to work as stated on the datasheet and if for whatever reason an antibody fails to perform as guaranteed, then we are happy to either replace or refund that antibody.

For ab7474, we guarantee it to work in IHC-P and IHC-Free Floating Sections on mouse and rat tissue, but we had not tested it in frozen sections, therefore that application is not covered under our Abpromise. However I would be very happy to help with any protocol suggestions, if you send me a copy of the protocol you are using with any variations you have tried.

Please let me know if there is anything else I can help you with.

I write you because I have a problem with yours antibody “Rabbit polyclonal to Oligodendrocyte Specific protein OSP” (aB7474) I ‘m making culture of primary glial cells and I want to discern oligodendrocytes from the others cellular types. I tried yours antibody with different dilutions (1/100, 1/50 and 1/25) to realize Immunoflorescent studies. The second antibodies that I use are: - anti-rabbit alexa fluor 488 (Invitrogen) (1/1000 for 2h) - anti-rabbit alexa fluor 555 (Invitrogen) (1/1000 for 2h) In every experiment that I have realized, I have obtained a poorly specific staining. (find enclose photos of the experiment performed with 1/25 dilution).

I would like to know if you give me some indications to optimize my immunofluorescent study.

ANSWER:

Thank you for your enquiry about ab7474. This antibody has not been tested in immunocytochemistry, though I would predict it has good chances of working in this application as it works very well in IHC on tissue sections.

I would like to suggest that we take a look in detail at your protocol to see what we can recommend to help improve the signal. Please find below a link to a protocol questionnaire to help you put your protocol details together easily; once we receive the form we aim to reply to you within 24 hours.

http://www.abcam.com/index.html?section=ihc&pageconfig=technical&intAbID=7474&mode=questionaire

Unfortunately we did not receive the photos you mention, if you can please attach those to the questionnaire these will also help us understand the problem.

If you would like some support in French, please put it to my attention; I am away from the office tomorrow but will gladly assist you on my return on Monday.

Does this Antibody recognize an extracellular epitope of OSP? Has it been used to stain cells for confocal microscopy?

ANSWER:

Thank you for your enquiry.

I am looking into whether this antibody recognizes an extracellular epitope. As soon as I have further information, I will let you know.

The antibody has not been tested with cells, but only in tissue sections. In addition, I do not have any information to suggest that the antibody has been tested in immunofluorescence.

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

Hi about the ab7474 OSP Ab, can this also blot for Claudin-11?

i would appreciate if you can answer my question.

ANSWER:

Thank you for your email. Oligodendrocyte Specific Protein OSP is a synonym for Claudin-11. So yes, ab7474 detects Claudin-11.

Please contact us again if you have any additional questions.

I purchased OSP ab (ab7474) and used for Western blots in an experiments where rat myelin/oligodendrocyte derivatives would be expected in the samples (but may be modified). The MW of the reactive band was ~68kD using 1:2000 dilution, quite distinct and specific.Gel was SDS reducing conditions. Do you have anymore information on yours or others investigations of CNS tissue? I noticed you state in the data that it reacts with 74kD band with CNS tissue. Was this gel run under reducing condtions? Do you know of others who have done CNS studies with the Ab (of any kind)?

ANSWER:

Thank you very much for your enquiry and your patience. I had to enquire with the originator of this antibody and have just heard back.

First, there was a mistake on the online datasheet for ab7474. It stated that the antibody detects a 74 kDa protein, but the originator told me that the antibody reacts with a 22 kDa protein from rat CNS samples. I have updated the datasheet to reflect this information and do apologize for this mistake.

The following reference used this antibody in Western blotting with mouse brain and testis samples: Hellani A et al. Developmental and hormonal regulation of the expression of oligodendrocyte-specific protein/claudin 11 in mouse testis. Endocrinology 141:3012-9 (2000). PubMed: 10919290 (the pubmed link is on the online datasheet for ab7474 and the full-text version is free).

The authors used a 15% SDS-polyacrylamide gel, used the antibody at 1:500 (diluted in TBS, incubated for 2 h at RT), and blocked with TBS 1%BSA.

I don't have any specific information regarding the use of ab7474 with rat CNS samples.

If you have any additional questions, please do let me know.