Anti-eIF4G1 抗体 (ab2609)

Thank you for your enquiry. The information on the datasheet was from a mail correspondence with the product originator back in June 2004 (please see FAQ section on datasheet). I am afraid that with our current mail system, we can not retrieve any old enquiries before the year 2006 therefore I am not able to provide more information on the enquiry itself. However, I did manage to do an internet search and found the following publication that mentions the susceptibility of eIF4G1 degradation. Caspase-3 is necessary and sufficient for cleavage of protein synthesis eukaryotic initiation factor 4G during apoptosis. FEBS Letters. Volume 451, Issue 3, 28 May 1999, Pages 332-336 http://linkinghub.elsevier.com/retrieve/pii/S0014579399006146 I am afraid that I can't provide any more information that already stated above but I hope that the publication and the FAQ information will be helpful.

Thank you for your enquiry. EIF4G is more susceptible to degradation compared to other proteins, especially from some tissue sources such as liver. This is true even when tissue is stored at frozen. SDS-PAGE sample buffer may improve the stability, but samples that are stored frozen may show degradation bands that have been described in the literature. Therefore if multiple bands are observed in WB, it is likely due to the degradation of eIF4G rather than non-specificity of the antibody. I would therefore recommend to take all precautions possible to prevent degradation of eIF4G (fresh protease inhibitors, keeping samples on ice at all times, using SDS-PAGE loading buffer), Please do not hesitate to contact us if you require further assistance,

I'm very sorry to hear you are having problems with a new vial of ab2609. Thank you for taking the time to fill in the questionnaire, it is very useful for me to understand your problem. I have checked the batches you have used and found out that they are actually derived from the same batch of the originator. Therefore it is likely that there was a problem with your particular vial, maybe it was damaged during transport or storage. I would like to send you a replacement vial immediately, could you please give me the Abcam order number or PO number and the name of the person who placed the order. I look forward to hearing from you,

As requested we will refund you for the cost of this antibody. I note from our datasheet that this antibody has only been tested for cross reactivity with human and rat and you advise that you were using Baby Hamster Kidney cells. However, the fact that other antibodies to eIF4G1 have worked successfully in your research means that there could also be a problem with the vial you received or with the quality of the product. Having checked other enquiries, I see there is another unfavourable report from a customer using this antibody that we received on Monday 28-June-2004 - please see the enquiries section of the online datasheet for further details. I will ask the originator of the antibody to re-test this product and if I am not satisfied with the result will remove it from our range. Our reputation is far too valuable to risk selling products that do not work as we say they should. Thank you for your feedback, and once again apologies for the inconvenience caused. [10 September 2004] We have confirmed the quality of this product with the originator. The problems this researcher experienced are most likely to have arisen because of the break down of eIF4G1 which is very unstable rather than problems with the quality of this antibody. eIF4G breaks down very quickly and it needs to be the first thing probed for when the test material comes out of the freezer. The shorter the time between Western blot and freeze clamping the tissues or freezing the cells the better. It is also work noting that I had misread the original enquiry from this researcher. The other primary antibodies used were raised against different proteins which would be more stable that eIF4G. We are confident that this antibody should remain in our range and would welcome feedback from other researchers using it.

Thank you for your patience. I'm sorry to hear that you are experiencing trouble with this antibody. I'm trying to obtain more information from the originator to help figure just what may be going on, but am still waiting to hear back. There is no need to send back the remaining antibody. I will send you another antibody as a replacement, or give you a refund. Can you clarify what you mean by there being no match between your IP samples and the controls? What size band did you see with the rat liver lysate?

Thank you for your patience. The originator of ab2609 has provided me with the following information: "eIF-4G migrates in SDS-PAGE just above myosin standard. If colored molecular weight markers are used it can be just above or just below dyed myosin standards depending on the manufacturer. EIF4G is more susceptible to degradation compared to other proteins, especially from some tissue sources such as liver. This is true even when tissue is stored at frozen. SDS-PAGE sample buffer may improve the stability, but samples that are stored frozen may show degradation bands that have been described in the literature." I don't know if you have done this already, but it is recommended to use rat liver lysate as a positive control. I have not received information regarding the IP buffers, but the originator has said that those in the literature should work fine. If you have any more questions or concerns, please do let me know.

The peptide used to generate the antibody is common to all 5 isoforms shown for SwissProt entry Q04637 (http://au.expasy.org/cgi-bin/niceprot.pl?Q04637). It lies between residues 560 and 660 of isoform A as defined in SwissProt entry Q04637.