Rabbit polyclonal Secondary Antibody to Rat IgG - H&L (HRP) (ab6734)

Regarding the antibody ab6734 (HRP conjugated rabbit polyclonal against rat IgG), please could you tell me roughly how many Horseradish Peroxidase (HRP) are there per antibody (is it 1 or 2 HRP per antibody?), and which conjugation method (is it biotin-avidin conjugation?) was used to attach the HRP to the antibody? Is there any cross-species reactivity?Is the 2mg/ml concentration of the vial the concentration of the HRP-conjugated polyclonal?

Same questions regarding antibody code ab7104.

Thank you for your help.

With best wishes,

ANSWER:

Thank you for your enquiry.

I have received a reply from the originator of ab6734 and ab7104. For both products, there are approximately two HRPs per antibody. The method we use for our HRP conjugation is proprietary. ab6734 is only tested against rat IgG and cross reactivity is possible, especially with species that are similar such as mouse and other small animals. Product ab7104 is preadsorbed against human serum proteins and shows no cross reactivity with human serum proteins. However, this may cross react with similar species as well as this has not been tested. Lastly, the concentrations listed on the data sheet are the concentration of the HRP conjugated antibody.

I hope this information helps. Please don't hesitate to contact us again if you have further questions.

Thanks for your response. So what is this integral membrane protein? Actually, the band I am getting is close to 25 kDa. So this is not TNF-alpha? Please tell me more about this integral membrane protein. I thought it is TNF-alpha because it is really a very clean band. Please advise me what dilution I need to use for the primary TNFantibody(ab11564) and secondary rabbit polyclonal to rat(ab6734).If you have a protocol please send me a copy. Thanks.

ANSWER:

Thank you for your email. Our source for this antibody tested ab11564 in Western blotting with recombinant TNF alpha. Conditions were standard. To use this antibody in Western blotting, I would recommend referring to our general Western blot protocol which can be found in the protocol section of the online datasheet. I would suggest using ab11564 at a 1:1000 dilution, and ab6734 at a 1:5000 dilution.

I conferred with our source for ab11564 and it sounds like you are detecting the membrane bound form of TNF alpha (the 26 kDa integral membrane protein).

If you have any additional questions, please contact us again.