Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090)

Thank you for contacting us.

The Goat anti-rabbit HPR Conjugated used in AB80436 has been developed specifically for use in this kit and is currently not available for separate purchase. We do offer a number of Goat anti-rabbit secondaries which are HRP conjugated and have been validated for used in IHC. AB7090 and AB98512 would each be excellent choices for any IHC against an antibody raised in rabbit. Each are pre-absorbed to reduce cross reactivity, and AB98512 is an F(ab')2 fragment. Use of F(ab')2 fragments further reduce non-specific background as these antibodies lack the Fc region which may bind endogenous Fc receptors within the tissue.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for your reply.

Reviewing the details, if you obtaining a band at 75 kDa in the no primary control with many secondary antibodies with the rat kidney lysate, but not the other samples, then I would recommend there is something in the rat kidney lysate that is crossreacting with these secondary antibodies.

It would be difficult to determine what the difference with the rat samples may be. However, one suggestion is thatendogenous rat antibody in the sample may be crossreacting with the secondary. Particularly in the case of infection or kidney disease, antibody could build up in the kidney tissue.

In this case:

1. The IgG antibody isotype has a molecular weight of 150 kDa which in reducing and denaturing conditions would break down to heavy and light chains and show on a blot at 25 kDa and 50Kda. However, if not fully reduced anddenatured,the heavy and light chain togethercould possiblyshow at 75 kDa. This may be what you are observing. I can suggest to ensure the sample is fully reduced and denatured when running on the gel.

2. I would suggest to try a pre or cross absorbed antibody to reduce the risk of crossreactivity with the rat IgG in the sample, but it seems you have already done so.

3. In case of problems with the individual rat, or possible contamination of the sample, could you confirm if you have tried running kidney samples from different rats?

I am very sorry there are no further explanations to provide in this case. Regrettably, it seems to be a samplephenomenon with the rat kidneyin this case.

If you have any further questions or concerns, please do not hesitate to contact me and I will be pleased to help.

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from these two secondaryantibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that both these antibodies aretested and covered by our 6 month guarantee for use in WB. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would appreciate if you can confirm some further details. It is very unusual to see this type of problem from several secondary antibodies. Please confirm:

1. The details you provideindicate several secondaries are giving a band at 75 kDa in the no primary control tests. Please confirm further details of any other secondary antibodies you are having this difficultywith.

I am concerned that something in the sample is crossreacting with the secondary antibodies:

2. Please confirm more details of how the samples are being prepared?

3. Is there anything special about these samples? Where the rats treated in any way? Or are the samples from just ordinary rat kidney from an untreated rat?

4. Have any other samples been tried, from a different species or tissue? Does this give the same result?

5. Have the membranes been stripped and reprobed at all? Or are they fresh membranes?

I hope this information is helpful, thank you for your cooperation. I look forward to receiving the requested details which will help us to investigate. I hope we can resolve this case as quickly as possible for you.

This antibody has been pre-adsorbed against Bovine, Chicken, Goat, Guinea Pig, Horse, Human, Mouse, Rat and Sheep Serum Proteins. This minimises the cross reactivity of this antibody with these proteins. However, minimal cross reactivity will still be seen. The non-adsorbed version of this antibody will therefore exhibit a greater cross reactivity to the above proteins than the pre-adsorbed. If you require an antibody towards rabbit IgG that does not react with mouse IgG, then we suggest a "Mouse polyclonal to rabbit IgG", as antibodies from the same species can't react with each other. Unfortunately, we don't stock this product at present.