EXPOSE Rabbit specific HRP/DAB detection IHC kit (ab80437)

IHC-P with mouse tissue
problem: no staining
AR; citrate buffer, heat (microwave)
Ab: 1/100 overnight
block: see kit ab80437
detection: kit ab80437
double staining with caspase 3 (rabbit poly ab13847)
send double staining protocols

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab56313.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I have attached the protocols for double staining.

I wish you the best of luck with your research. If you have further questions, please let me know.

Hi,

A customer is enquiring about the use of “EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit” with a primary antibody which is raised in rat (rat anti mouse CD144 - IgG1). He wants to know if there is any way he can use the Expose kit? I suggested that the only way is to use an extra step ie a rabbit anti rat secondary followed by the goat anti rabbit-HRP (from the expose kit). Has anyone done this before? Is there any reason why this would not work. In any case, he does not have any rabbit anti rat on hand – if that would work, can you suggest a suitable one he could buy?

Thanks.  Kind regards,   

ANSWER:

Thank you for contacting us.

The EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit (ab80436) can be used to detect mouse and rabbit primary antibodies. You are therefore correct in thinking that the kit would need to be modified in order to detect a Rat primary antibody. This could potentially be done by using a different secondary antibody to "change the species" or the primary antibody, although we do not have any experimental data regarding this, in theory it should work.

The concentration of the rabbit anti-rat antibody would need to be optimized by the user, I would recommend they try ~10 ug/ml to start. I would also suggest that the customer should purchase the ab80437 kit instead of the ab80436 kit, since they do not require the rabbit anti-mouse antibody for their application.

When choosing the antibody to use your customer has to be a bit careful as the kit only detects antibodies of the IgG isotype. An antibody that could be used for this application is ab102205 which is a rabbit polyclonal (IgG) to rat IgG. I hope this information has been of help.

If you require any further information please do not hesitate to ask.

How long is this kit stable for?

ANSWER:

Thank you for calling Abcam earlier today.

I have had a response from the lab about the storage life of the kit, ab80437, the EXPOSE Rabbit specific HRP/DAB detection IHC kit. They have told me that the storage life of this kit is 12-18 months if stored as recommended at 2-8C and the actual expiration date for each product should be indicated on the label.