Anti-ICAM1 antibody [MEM-111] (ab2213)

This is your ref:ORDER xxx
The product was purchased the xxx

ANSWER:

Thank you for your reply.

The options of resolving this case are to replace the antibody free of charge with another antibody (different lot; same target; different target)or to issue a credit or refund.

I assume from the email conversation with my colleague that you have used human tissue?

We carry several ICAM1-antibodies that have been tested in human and IHC-P and from which you can choose from:
ab53013, ab67217, ab109361, ab20144, ab67345and ab89378.

I look forward to hear back from you and resolve this case.

The stromal staining is considered to be correct staining in the
appendix because I'm looking on the
http://www.proteinatlas.org/ and the stromal staining in the
appendix is considered to be a positive control.
I have used the protocol of the data sheet:
Tissue was fixed with formaldehyde and an antigen retrieval step
was performed using EDTA, pH 8.0, 20 minutes at 100°C. Samples
were then incubated with ab2213 at a 1/50 dilution for 1h at 25°C.
The secondary used was a DAB detection system.

ANSWER:

Thank you for your reply and the extra information.

Your protocol looks fine to me. It might be the antibody that is the problem then.

Could you please let me know your order or PO number?

If the product was purchased within the last 6 months, I'd be happy to replace, credit or refund theproduct.

I look forward to hear back from you and resolve this case.

I am contacting you for a technical problem. I tested your ICAM-1 antibody (# ab2213, in immunohistochemistry.
I used lung and appendix tissues as a positive control.
The labeling is very beautiful in the appendix, I thing my protocol is good. However, I have no label in the lung and labeling in the appendix does not fit the theory (marking the stroma between the glands). Here I have glands that are marked as if it were a marker of epithelial cells and did not mark the stroma.
I have put my pictures of immunohistochemistry in attachment.
Have you had similar remarks or can you help me regarding this problem?
Thank you in advance,

ANSWER:

Thank you for sending your IHC images.

Is the stromal staining considered to be correct staining in the appendix? I have not seen many examples of ICAM1 staining but my understanding is that it is expressed in endothelium and leukocytes. The examples on the datasheet are not good images. The stain is strong but I am not convinced it is correct.

In any case, your lung tissue stain is weak or non-existent, suggesting the antibody is defective. We have not received similar complaints, though I do see one about non-specific staining and high background.

Can you please send me your protocol? I understand that you think it is good, but there may be something I can suggest to improve the results. If not, I will be happy to discuss a replacement or refund. Please also send your order information (the PO number or date of order).

Has used a full vial while troubleshooting to reduce background in IHC on human FFPE tissue.

ANSWER:

Thank you for your call today and for letting us know about the trouble with ab2213.

As we discussed, I'm sending a free of charge vial on the order *** which should arrive tomorrow.

Please keep me updated about any further progress with this antibody, and let me know if you need anything else. Have a great day!

Hi, I would appreciate it if you can provide tech support on this antibody. See attachment for details.

ANSWER:

Thank you for contacting Abcam.

I am sorry that you are having problems with ab2213 in western blot. Under our Abpromise, we do guarantee it to work on human tissue in western blot for 6 months. If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

I just had a few more protocol questions to ask you:

1 - What primary antibody concentration have you tried? We recommend starting at 1/400.

2 - What size are the non-specific bands that you are seeing? Or are there no bands present on the membrane?

3 - How much total protein (in ug) did you load?

I look forward to your reply and helping you resolve this issue.