Anti-MAP2 antibody - Neuronal Marker (ab32454)

My customer tried the MAP2 antibody again, with no luck. Please see attached protocol. She also had some problems with AB290 (PO 137874), can you please take a look at that protocol as well?

ANSWER:

Thank you for contacting us.

I am sorry that the protocol recommendations that were suggested did not work. We can replace the anti-Map2 antibody with another vial of the same product or with another product. I would suggest the rabbit polyclonal ab70218 (http://www.abcam.com/MAP2-antibody-ab70218.html) orone the mouse monoclonal antibodies ab11268 (http://www.abcam.com/MAP2-antibody-AP-20-Neuronal-Marker-ab11268.html) or ab11276 (http://www.abcam.com/MAP2-antibody-HM-2-Neuronal-Marker-ab11267.html). These have all been tested in IHC-P and are covered by our Abpromise guarantee.

As to the difficulties with ab290, this one of our most characterized antibodies. I would suggest trying a heat mediated antigen retrieval step with citrate buffer pH 6.0.


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

I had a customer contact me regarding a problem with background using AB32454.  Please see protocol details attached.  This antibody was ordered on.   Thank you for looking into this for me.

ANSWER:

Thank you for contacting Abcam.

I am sorry to hear of your customer's difficulties when using this product and wish to thank them for providing me with their protocol information. While reviewing this I did happen to notice that the perform and antigen retrieval step as well as permeablization. While this in itself should cause no issues, what is problematic is that the permeablization step is so harsh, the 1% tween used is 10 times higher than we recommend, that I am afraid this plus the retrieval step is greatly affecting their tissue. I would recommend that future experiments do not include a permeablization step. There are many Abreviews on the webpage for this product where other customers have not used permeablization and have achieved excellent results.

I would suggest changing the blocking conditions used with this product. Others have had good success using a BSA blocking buffer.

I would also suggest a reduction the incubation time of the primary antibody. If these samples have been incubated at 4C overnight, it might be good to try 2 hours at room temperature or less time if the incubation is at room temperature now.

Please let me know the results of any further testing as well as any questions or concerns that may arise.