Anti-L Glutamate antibody (ab9440)

Thank you for contacting us.
Here are the references we spoke about on the phone for this product: https://www.abcam.com/L-Glutamate-antibody-ab9440-references.html
Some other background references might be helpful as well to see what other labs have seen for staining.
http://www.ncbi.nlm.nih.gov/pubmed/15215299
http://www.ncbi.nlm.nih.gov/pubmed/18067143
http://cercor.oxfordjournals.org/content/10/11/1132.full
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for contacting us and for your interest in our products. I can confirm that ab9440L Glutamate antibody is tested and covered by our guarantee for IHC-FoFr, IHC-Fr, ICC, Immunomicroscopy andIHC-P. I am sorry it has not been tested in ELISA or western blot. All tested applications are listed on the datasheet, and these are updated as soon as any new information is brought to our attention. With regards to publications, the following 2 publications have used the antibody in IHC: Sun D et al. Metabolic and functional profiling of the normal rat retina. J Comp Neurol 505:92-113 (2007). IHC; Rat. PubMed: 17729258 Wilkie IC et al. Physiological and immunocytochemical evidence that glutamatergic neurotransmission is involved in the activation of arm autotomy in the featherstar Antedon mediterranea (Echinodermata: Crinoidea). J Exp Biol 213:2104-15 (2010). IHC-R; Starfish. PubMed: 20511525 I am sorry we have no other L GLutamate antibodies tested in rat that are also tested in WB and ELISA. However, if you would like to test the ab9440 antibody in ELISA or Western blot application, please contact me again prior to the purchase by replying to this message as you may be eligible for our testing discount program. Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates. To find out more about our Abreview system, please see the following link: https://www.abcam.com/abreviews I hope this information is helpful. Please do not hesitate to contact me for any further advice or information.

Thanks for your call today and for your patience while I have looked into your question. I found that the antibody ab8893 does not react with free L-glutamate so I don't believe it will be suitable for the ELISA assay that you are planning. We do have a couple of L-glutamate antibodies, ab9440 and ab62668, that do not react with glutamate in peptides or proteins, so it will be specific for the free amino acid. However, we have not tested these antibodies in ELISA. We do have a testing discount program if you would like to try either of them in ELISA, and I would be happy to send more information about this program if you would like. Please let me know if you have any questions, or if there is anything else that we can do for you.

I have received some further information regarding this antibody from the priginator. Ab9440 does not bind any anti-mouse product that has been tested. The only possible explanation is that the antibody was denatured and that it exposed a non-specific heavy chain motif that the anti-mouse IgG likes, but disrupted the anti-rabbit IgG motif. Any tissue binding was likely non-specific. Some protocol notes that I received - Ab9440 was explicitly designed for glutaraldehyde (GA) fixation and plastic-section detection, especially ultrastructure (e.g. Ottersen 1989 Postembedding Immunogold Labeling of Fixed Glutamate - an Electron-Microscopic Analysis of the Relationship between Gold Particle Density and Antigen Concentration Journal Of Chemical Neuroanatomy 2:57-66). OPTIMAL FIXATION: 0.1-2.5% glutaraldehyde,1% formaldehyde using HPI protocol (please see the protocols tabe on the online datasheet). The residual free glutamate can be detected with FA if the protocol is optimized (>48h fixation, proper temp, pH), yielding enough time for slow methine-bridge formation. See Hayat 1981 Fixation for electron microscopy; Hayat 2001 Principles and Techniques of Electron Microscopy: Biological Applications). If fixation is brief, most or all of the glutamate will be lost. Without GA fixation, over 95% of all amino acids will be lost. Glutamate signal strength will decrease nearly 2 log units. If cells are loaded with 3H-glutamate and fixed with FA, virtually all the free 3H-glutamate is lost. In addition, it is recommend to use a positive tissue control with GA fixation and resin sections - retina, brain, spinal cord will all have strong signals. As I mentioned in my previous email, I can send you a different batch to try. Please let me know if you would like to try it and please send me the Abcam order number or purchase order number that was used when you originally purchased ab9440.

Thank you for your enquiry. Unfortunately, we are not aware of any publications that feature the use of this antibody. When we are aware of such references, we cite them on the online datasheets. If you have any more questions, please contact us again.