Overview

  • Product name
  • Description
    Rabbit polyclonal to RAGE
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, Flow Cyt, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Synthetic peptide:

    KGAPKKPPQRLEWKLNTGRTC

    , corresponding to amino acids 39-58 of Human RAGE

Properties

Applications

Our Abpromise guarantee covers the use of ab37647 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
ICC/IF Use a concentration of 1 µg/ml.
WB 1/1000 - 1/5000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
Flow Cyt Use 1µg for 106 cells.

Use PBS/EDTA to detach cells to preserve the glycoproteins on the cell surface; do not fix, do not permeabilise.

 

 

 

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

 

ELISA 1/2000 - 1/10000.

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
    • MGC2235 antibody
    • RAGE_HUMAN antibody
    • Receptor for advanced glycosylation end products antibody
    see all

Images

  • All lanes : Anti-RAGE antibody (ab37647) at 1/500 dilution

    Lane 1 : HUVEC whole cell lysate
    Lane 2 : EVC304 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP conjugated goat anti-rabbit

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 17,40 kDa
    why is the actual band size different from the predicted?


    Exposure time: 4 minutes


    The band at 52kDa is a tubulin loading control.

    See Abreview

  • RAW 264.7 cells were stained with anti-RAGE (1ug/million cells) for 1hr at 4 deg C followed by staining with anti-rabbit IgG-PE conjugate. Cells were analyzed by flow cytometry. Unstained cells or cells stained with secondary antibody alone are represented in the background
  • ab37647 at a 1/100 dilution staining RAGE in bovine endothelial cells by Immunocytochemistry/ Immunofluorescence. Fixed in PFA, permeabilized with Triton X-100. Blocked using 10% serum for 20 minutes at room temperature. Secondary used at 1/200 polyclonal Goat snti-rabbit conjugated to Alexa Fluor 568 (red). Nuclear staining (blue).

    See Abreview

  • All lanes : Anti-RAGE antibody (ab37647) at 1/2000 dilution

    Lane 1 : Bovine lung extract
    Lane 2 : Mouse lung extract
    Lane 3 : HeLa cells expressing human soluble RAGE

    Secondary
    All lanes : anti-rabbit IgG alkaline phosphatase conjugate

    Predicted band size: 42 kDa
    Observed band size: 42 kDa



    Bovine RAGE is slightly larger than human and mouse RAGE - 44kDa compared with 42kDa
  • ab37647 staining human Jurkat T cells by Flow Cytometry. The cells were prepared in PBS with 0.2% BSA. The primary antibody diluted 1/100 and incubated with sample for 30 minutes at 0°C. The secondary antibody was Alexa Fluor® 488 conjugated goat polyclonal to rabbit IgG, diluted 1/200.

    Specimen tube 006 is negative control

    See Abreview

  • ICC/IF image of ab37647 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab37647, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Wu DM  et al. S100A9 gene silencing inhibits the release of pro-inflammatory cytokines by blocking the IL-17 signalling pathway in mice with acute pancreatitis. J Cell Mol Med 22:2378-2389 (2018). WB ; Mouse . Read more (PubMed: 29441717) »
  • Jandial R  et al. Inhibition of GLO1 in Glioblastoma Multiforme Increases DNA-AGEs, Stimulates RAGE Expression, and Inhibits Brain Tumor Growth in Orthotopic Mouse Models. Int J Mol Sci 19:N/A (2018). Read more (PubMed: 29385725) »
See all 32 Publications for this product

Customer reviews and Q&As

1-10 of 22 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Brain, temporal cortex)
Antigen retrieval step
Other
Permeabilization
Yes - hydrogen peroxide in methanol
Specification
Brain, temporal cortex
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1.5% · Temperature: RT°C
Fixative
Formaldehyde

Miss. Annabelle L. Chambers

Verified customer

Submitted Oct 11 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (Human gingival epithelial cells)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
20 µg
Specification
Human gingival epithelial cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 21 2015

Application
Western blot
Sample
Mouse Tissue lysate - whole (gingival tissue)
Gel Running Conditions
Reduced Denaturing (4-15% GRADIENT GEL)
Loading amount
20 µg
Specification
gingival tissue
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 01 2015

Answer

Thank you for your enquiry.

I can confirm, as stated on the datasheet, that the immunogen for ab37647 Anti-RAGE antibody is as follows:

KGAPKKPPQRLEWKLNTGRT

This is corresponding to amino acids 39-58 of Human RAGE

I am sorry to confirm that this has not been specifically tested for detection of the different isoforms. However, I have reviewed the sequences for you. According to the Uniprot link on the datasheet for the human protein, the immunogen sequence is contained in the following isoforms:

Isoform 1, mwt 42,803
Isoform 3, mwt 37,050
Isform 4, mwt 39,020

Isoform 2 only contains half the immunogne sequence, and so the antibody probably won't cross react with this isoform (mwt 36,193).

I hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer


With the two antibodies you have been looking at I would suggest the following antibodies should serve as suitable isotype controls:

Rabbit polyclonal to RAGE (ab37647), rabbit IgG, isotype control: Rabbit IgG- ChIP Grade (ab37415)

Mouse monoclonal to RAGE (ab54741), mouse IgG2a, isotype control: Mouse Control IgG2a [MOPC-173] - Chip Grade (ab18413)

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (stromal cell)
Specification
stromal cell
Fixative
Formaldehyde
Permeabilization
Yes - Saponin
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Apr 02 2013

Answer

Merci de nous avoir contactés.

Nous sommes désolés d'apprendre que le produit que vous avez reçu ne fonctionne pas comme attendu. Je me permets de vous transférer le numéro de commande de remplacement gratuit de ce produit : xxxxx. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition. Normalement, vous devriez le recevoir jeudi.

N'hésitez pas à nous contacter lors d'une prochaine occasion.

Read More

Answer

Merci d'avoir pour ce supplément d’informations.

Malheureusement nous n’avons pas d’autre lot en stock pour ab37647 et je crains que si je vous envoie en remplacement le même lot, que vous obtiendrez les mêmes résultats.

Je peux vous proposez ab3611 (https://www.abcam.com/ab3611) qui est aussi garanti de fonctionner en ICC/IF et WB chez le rat. Sinon, je peux également vous faire un avoir ou un remboursement.

J’attends votre réponse en ce qui concerne la façon dont vous voulez procéder avec cette affaire.

Read More

Answer

Thank you for contacting us.

We have not tried this antibody in functional assay using living cells so I am sorry we are unable to confirm how well this antibody will perform. There are two basic things to consider when using ab in vivo, the preservative and endotoxin level. This antibody does have 0.1% Sodium Azide which is known to be a cytotoxic agent so it makes this product unsusable however the effect might be minimal at high dilution of use. We have also not determined the endotoxin level of this antibody which is another reason why we will not be recommending it.

The ab37647 is a good candidate which has been used in neutralization application, you might use this ab in vivo.

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

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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1-10 of 22 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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