Overview

  • Product name

    Anti-RAGE antibody [MM0520-8D11]
    See all RAGE primary antibodies
  • Description

    Mouse monoclonal [MM0520-8D11] to RAGE
  • Host species

    Mouse
  • Tested applications

    Suitable for: Flow Cyt, WB, Neutralisingmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant Human RAGE extracellular domain.

Properties

Applications

Our Abpromise guarantee covers the use of ab89911 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use at an assay dependent concentration. PubMed: 24567328

ab18428 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 

WB 1/500 - 1/1000. Predicted molecular weight: 48 kDa.
Neutralising Use at an assay dependent concentration.

Target

  • Function

    Mediates interactions of advanced glycosylation end products (AGE). These are nonenzymatically glycosylated proteins which accumulate in vascular tissue in aging and at an accelerated rate in diabetes. Acts as a mediator of both acute and chronic vascular inflammation in conditions such as atherosclerosis and in particular as a complication of diabetes. AGE/RAGE signaling plays an important role in regulating the production/expression of TNF-alpha, oxidative stress, and endothelial dysfunction in type 2 diabetes. Interaction with S100A12 on endothelium, mononuclear phagocytes, and lymphocytes triggers cellular activation, with generation of key proinflammatory mediators. Interaction with S100B after myocardial infarction may play a role in myocyte apoptosis by activating ERK1/2 and p53/TP53 signaling (By similarity). Receptor for amyloid beta peptide. Contributes to the translocation of amyloid-beta peptide (ABPP) across the cell membrane from the extracellular to the intracellular space in cortical neurons. ABPP-initiated RAGE signaling, especially stimulation of p38 mitogen-activated protein kinase (MAPK), has the capacity to drive a transport system delivering ABPP as a complex with RAGE to the intraneuronal space.
  • Tissue specificity

    Endothelial cells.
  • Sequence similarities

    Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Cellular localization

    Secreted and Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • Advanced glycosylation end product-specific receptor antibody
    • Ager antibody
    • DAMA 358M23.4 antibody
    • MGC2235 antibody
    • MGC22357 antibody
    • RAGE_HUMAN antibody
    • Receptor for advanced glycation end products antibody
    • Receptor for advanced glycosylation end products antibody
    see all

Images

  • Flow Cytometry analysis of human mesenchymal stem cells labelling RAGE with ab89911. A Phycoerythrin-conjugated F(ab′)2 goat anti-mouse IgG was used as secondary antibody. Histograms were set according to negative control stainings (#).

References

This product has been referenced in:

  • Jang EJ  et al. HMGB1 enhances AGE-mediated VSMC proliferation via an increase in 5-LO-linked RAGE expression. Vascul Pharmacol N/A:N/A (2019). Read more (PubMed: 30954689) »
  • Huang CY  et al. HMGB1 promotes ERK-mediated mitochondrial Drp1 phosphorylation for chemoresistance through RAGE in colorectal cancer. Cell Death Dis 9:1004 (2018). Read more (PubMed: 30258050) »
See all 5 Publications for this product

Customer reviews and Q&As

1-6 of 6 Q&A

Answer

Thank you for your phone call. I'd be happy to issue you either a credit or refund. Alternatively, a free of charge replacement with another antibody can be sent. I have checked our catalog for other potential suitable RAGE antibodies to be used in neutralization/receptor inhibition. As I am not certain where in the extracellular domain (residues 23 – 342) the AGE binding site would be, I have selected all RAGE antibodies with immunogen in the extracellular domain: ab37647 (https://www.abcam.com/RAGE-antibody-ab37647.html) ab7764 (https://www.abcam.com/RAGE-antibody-ab7764.html) ab78022 (https://www.abcam.com/RAGE-antibody-DD-A11-or-mAbA11-ab78022.html) ab86767 (https://www.abcam.com/RAGE-antibody-ab86767.html) ab87305 (https://www.abcam.com/RAGE-antibody-Azide-free-ab87305.html) None of these antibodies was tested in neutralization and would therefore qualify for our testing discount program. For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works: The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. Please let me know how you would like to proceed and I'd be happy to arrange it for you. I look forward to hear back from you.

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Answer

Thank you for your patience. The lab send the follwoing information and suggestion: (1) The immunogen of this antibody is the extracellular domain (based on the UniProt database entry Q15109: residues 24 to 344). (2) The customer could determine whether this antibody is agonistic by measuring TNFa in cell culture media when ab89911 is added alone (without AGE-BSA). We'd suggest to try this. I am not sure if you have done this already, but maybe it would help to measure TNF-a under the following conditions: ab89911 alone AGE-BSA alone ab89911 + AGE-BSA together Titrate ab89911 and AGE-BSA separately as well as together. You might want to use similar concentrations as the lab did for their ELISA to make sure you see the blocking effect (2 μg/mL ab89911, 0.5 μg/mL of Biotinylated-AGE-BSA). How much RAGE do you think is expressed on the PBMCs? Should after trying these tips the results still not be satisfactorily, I'd be happy to replace, credit or refund the antibody. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us. I have passed on your experimental details to the lab for more input, and will let you know what they suggest. Regarding your questions: 1) Which part of the epitope is this antibody raised against? The antibody was raised against the extracellular domain which is about 300 residues long. The exact epitope (which is usually 5-10 residues) has however not been mapped. 2) Is there an available full length anti-RAGE that blocks it and inactivates the AGE-RAGE pathway? I am not sure if I understand this question correctly. AGE would be binding to the extracellular domain only. Thus an antibody against full length RAGE would not make much difference or even block binding. We do not carry any other antibody that has been tested so far in inhibition studies - only ab89911. Lets wait if the lab has any further suggestions and we can discuss further how to proceed. I will be in touch. In the meantime, please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us. Here is the reply I received from the lab. First to answer your question about the testing protocol used by the lab for neutralization studies: The neutralization function of this antibody was tested in vitro using a functional ELISA (between two proteins on ELISAs plates): Biotinylated AGE-BSA can bind to immobilized recombinant human RAGE coated on an ELISA plate. When 4 μg/mL of RAGE (100ul) was coated on to the ELISA plate, a 2μg/mL of ab89911 can block 50% of the binding of 0.5μg/mL of Biotinylated-AGE-BSA to RAGE and at 10 μg/mL, this antibody can block 92% of the binding. (The suggested conditions are for reference only and the end users should optimize the conditions themselves). Second, some suggestions for your experiment: Our test indicates that ab89911 can block the binding between AGE and RAGE. In your experiment, no AGE is present in the system (right?). We are not sure what you should expect in this study by simply using ab89911 to block RAGE. We suggest that the readout would need to be specific for the downstream signal after AGE-RAGE binding for this blocking experiment when AGE is added into the system. Also, I am not really sure why the TNF alpha signal would increase when using this antibody. Thus, I was wondering if maybe TNF alpha is not the right signal to measure? Or maybe could the antibody cause an immune response with the PBMCs and thus increase the TNF alpha signal? I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thanks for your inquiry. Our neutralization assay was done in vitro between RAGE and AGE-BAS. It means the antibody indeed blocks the binding of the receptor from AGE in the in vitro setting. I hope this answers your question. Please feel free to contact us if you require further assistance.

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Question
Answer

Thank you for your calls yesterday, and for your patience. According to the lab, the antibody was tested in vitro using a functional ELISA as follows:   If uninhibited, biotinylated AGE-BSA can bind to immobilized recombinant human RAGE coated on Elisa plate. For the neutralization assay, 4 μg/mL of RAGE (100ul) was coated on to the Elisa plate, and 2μg/mL of ab89911 blocked 50% of the binding of 0.5μg/mL of biotinylated AGE-BSA to RAGE. At 10 μg/mL, this antibody can block 92% of the binding. We do not have any published references using this assay. I hope this information is helpful, but please let me know if you have any further questions.

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