Anti-eIF4E antibody (ab1126)

Thank you for your email. The issue for AB1126 is for the same customer. This started from August 2011.

This customer did receive a replacement of 1 vial form the same Lot# but initially he ordered 2, and this also failed for him.

I hope to hear from you soon.



Kind regards,

ANSWER:

Thank you for your response.
This is to let you know that I have just contacted and asked our Account Department to raise a credit note for you - for the cost of two vials of ab1126. For your information, the internal reference note for this credit is CN19459. You can use this credit in the near future for any of the products which are in the catalogue.
I hope this helps and if I can assist further, please do not hesitate to contact me.

Dear Technical team

Good day! We have a customer who has purchased the above antibody and would like to know the following information: As your website recommended the tissue and dilution of antibody I have done work like that.

Process:

Antibody dil- 1/1000

Breast Cancer Tissue

Mild and Standard Retrieval time

Automated Benchmark machine for imunostaining

Result:

With standard Retrieval time- strong peri nuclear staining but non specific with cytoplasmic as well.

With Mild retrieval time- Not specific staining.

Please check with my questions:

1. What kind of breast cancer tissue is suitable for this antibody as control?

2. Which retrieval time is suitable?

3. Which antibody dilution is suitable?

4. Is it tested on other cancer tissue?

5. What is the actual peri nuclear staining?

  I hope to hear from you soon. Kind regards

ANSWER:

Thank you for contacting us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

As it can be seen from the image in the datasheet, human breast carcinoma is a good option for a positive control. However, there are other cancer tissues that may also give successful results as positive control, these are: colorectal cancer tissue and prostate cancer tissue, you can find the information at the Human Atlas link:

http://www.proteinatlas.org/ENSG00000151247

Regarding the retrieval time, it really has to be optimised for each case. The standard recommended time is 20 minutes, less than 20 minutes may leave the antigens under retrieved, leading to weak staining. More than 20 minutes may leave them over-retrieved, leading to nonspecific background staining and also increasing the chances of sections dissociating from the slides. A control experiment is recommended beforehand, where slides of the same tissue section are retrieved for 5, 10, 15, 20, 25 and 30 minutes before being immunohistochemically stained to evaluate optimum antigen retrieval time for the particular antibody being used. In the following link you will find useful information for the different antigen retrieval methods:

http://www.abcam.com/index.html?pageconfig=resource&rid=11488#notes2

The optimal dilution for the primary antibody needs also to be optimised for every experiment. We suggest using 1/1000 as a starting point, and depending on the results you may need to use different dilutions.

All the information about this product is on the datasheet, we will be glad to update it with new data from customers. All new images and publications are welcome in order to provide more information to researches.

I would be very happy to help you with your experiments, but in order to do it I will appreciate to gather more information. I send you a questionnaire that only takes about 5 minutes to be filled. Any images will also be very helpful to understand why this antibody did not perform as stated in the datasheet.

All of our products are covered by our Abpromise guarantee. In case they fail to perform as expected, we will offer a free of charge replacement, a reimbursement or a credit note.

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

Dear Technical,   Good day! I was wondering if you could give us a recommended dilution for IHC-P for this antibody. As the recommended dilution states to use 1ug/ml but it is a bit difficult when the concentration of the antibody is not recommended. Could you please help me recommend a dilution for a customer?   I hope to hear from you soon.   Kind regards  

ANSWER:

Thank you for your enquiry regarding ab1126.

I can confirm that the antibody has been tested in IHC-P at a dilution of 1/1000 (instead of 1ug/ml). The datasheet (application notes) and the legend of the IHC-P image has been amended accordingly.

I apologize for any inconvenience caused and thank you for bringing this matter to our attention.

I have purchased a number of antibodies from Abcam and they have all been good. I have couple of questions which i would like you to assist me with, as i have checked your website and found no answers. The Rabbit polyclonal eIF4E antibody (ab1126) does it detect total eIF4E ie the non phospho and the phospho or just the non phospho form The Mouse Monoclonal eIF2a antibody (ab5369) does it detect total eIF2a ie the non phospho and the phospo or just the non phospho form

ANSWER:

Thank you for your enquiry.

I can confirm that both these eIF2a antibodies (ab1126, ab5369)will detect both phosphorylated and de-phosphorylated forms of the protein.

I hope this information is helpful. Should you have any further questions, please do not hesitate to contact us again.

Thanks for your rapid reply, it was a misunderstandig. Of course we know, that eIF4E is well expressed in HeLa. and we used for eIF4E a little aliquot of an ab which we got as a present from another lab. So previously we could detect eIF4E. The point is your ab results in a completly black western, even when I expose the film only for 5 sec. unfortunately we don't have any bovine kidney extract to test that your ab. regards

ANSWER:

Thank you very much for your e-mail.

I would like to reassure you that if an antibody does not perform as specified on the datasheet, we will provide a full refund or offer you a replacement of equivalent value.

I have contacted the supplier regarding your problem and will get back to you as soon as they reply.

We apologize for any inconvenience caused and thank you for bringing this matter to our attention.