Goat Anti-Rabbit IgG H&L (DyLight® 550) (ab96884)

My colleague forwarded your message about testing an Abcam IGF2 antibody to me.

As he mentioned, if western blotting and IHC are listed as applications on the datasheet, and rat and mouse are listed, then the antibody will be guaranteed for those applications and species and a replacement or credit or refund is available in the event it fails. If not, it is not guaranteed.

For untested applications or species, we do not have samples, but we offer the testing discount he mentioned.

The procedure is as follows: I first issue a discount code to you. Then, your lab purchases the antibody, tests it, and sends us your results in the form of a review (www.abcam.com/abreviews has some information about this). Once we have your review, your testing discount code becomes active, regardless of whether the results are positive or negative.

The discount is a credit that can be applied to a subsequent order, good for one free primary antibody of your choice. I will be happy to issue you a code before you purchase. I just need the antibody catalogue number, and I will send you a code, which you include in the notes section of the review form.

For some examples of customer reviews, please see the datasheet for anti-IGF2 ab9574:

Click here (or use the following: https://www.abcam.com/IGF2-antibody-ab9574.html).

Regarding a control peptide, this is available for some antibodies, but not all. If so, it will be a separate item for purchase.

I look forward to your reply.

I've been following up on the issues that we spoke about earlier in regards to the Anti-E Cadherin (phospho S838 + S840) antibody. I have also received your corresponence and thank you for taking the time to send that to me. I'll try to answer all the questions that you have raised and try to get these antibodies working for you. "1. Could primary Ab be nonspecifically attached to plastic in 96 well plate, causing high background?" Although there is a possibility your findings that "Minimal difference between groups 1 and 2 – cells with or without primary antibodies, possibly because of high background intensity" leads me to believe that background problems are a result of the secondary antibody.   "2. How to reduce the intensity of background: extra wash after introducing  primary Ab, going beyond ABcam recommended dilution rate for the secondary Ab?" I would definitly recommend decreasing the concentration of the secondary antibody as well as blocking in 5% BSA for 1 hour at room temp.  "3. Should we try to use higher concentration of primary Ab to possibly engage more e-cadherin sites for secondary Ab to differentiate from the background? Could that also increase the background?" This may help I would recommend doing this if you still have target recognition issues but only after you have been able to reduce your background staining. "4. Do we really need to conduct the cell fixation, and could that cause the cell loss in the experiment?" We spoke of the need to perform fixation to preserve protein morpholoy and expose epitopes. It is very possible that changing the fixation to acetone or methanol will greatly help you experiments. "5. Could cell permealization add to the amount of reacting epitopes?" I've found that the epitope site recognized by this antibody is intracelluar. Permeablization, whether by a detergents such as NP-40 or Triton-X 100 is neccessary if the fixative does not permeablize (eg , formalin, pfa), the acetone fixation that we spoke of will permeablize the cells so a seperate step will no be needed. I've included a pdf detailing ICC fixation and permeablization steps for you (see attached). Here is the SwissProt page where I found that information: http://www.uniprot.org/uniprot/P12830 "6. Conducting the experiment on  glass to prevent possible non-specific secondary Ab  attachment  to plastic causing high background? Could that also contribute to the higher cell loss after fixation? Could that possibly improve the resolution?" Although glass may help , I would definitely work with changing other aspect of the protocol first. Here is a link to our standard ICC protocol as I have not found a product specific one: https://www.abcam.com/index.html?pageconfig=resource&rid=12129 I hope that these suggestions help. Please feel free to contact us with any other questions that you have.