Overview

  • Product name
  • Description
    Mouse monoclonal to RAGE
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein (GST-tag) corresponding to Human RAGE aa 23-405. Full length mature chain, without signal peptide.
    Database link: Q15109

  • Positive control
    • WB: Human kidney tissue lysate. RAGE transfected HEK-293T cell lysate. Recombinant RAGE protein. Flow Cytometry: Jurkat cells. IHC-P: Human lung tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab54741 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
WB Use a concentration of 1 - 5 µg/ml. This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein in Western blot
IP Use at an assay dependent concentration.
Flow Cyt 1/100.

See Abreview.

 

 

 

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

  • Anti-RAGE antibody (ab54741) at 1 µg/ml + Human kidney tissue lysate

    Developed using the ECL technique.
  • ab54741 (1 µg/ml) staining RAGE in human lung using an automated system (DAKO Autostainer Plus). Using this protocol there is membrane staining throughout the alveloli.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

  • All lanes : Anti-RAGE antibody (ab54741) at 1 µg/ml

    Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) RAGE transfected cell lysate
    Lane 2 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) non-transfected cell lysate

    Developed using the ECL technique.
  • ab54741 staining human Jurkat (human T cell leukemia cell line from peripheral blood) 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 mouse IgG, diluted 1/200.

    Specimen tube 001 is negative control.

    See Abreview

  • Anti-RAGE antibody (ab54741) at 1 µg/ml + tagged recombinant protein immunogen

    Developed using the ECL technique.


    Predicted band size of immunogen is 68 kDa

  • Immunoprecipitation of RAGE transfected lysate using ab54741 and Protein A Magnetic Bead. Immunoblotted with a rabbit polyclonal anti-RAGE antibody.

References

This product has been referenced in:
  • de Carvalho GC  et al. Up-regulation of HMGB1 and TLR4 in skin lesions of lichen planus. Arch Dermatol Res N/A:N/A (2018). Read more (PubMed: 29728859) »
  • Lei H  et al. Human S100A7 Induces Mature Interleukin1a Expression by RAGE-p38 MAPK-Calpain1 Pathway in Psoriasis. PLoS One 12:e0169788 (2017). WB ; Human . Read more (PubMed: 28060905) »
See all 11 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Bel-7402 cells)
Gel Running Conditions
Non-reduced Denaturing (10% SDS-PAGE)
Loading amount
50 µg
Specification
Bel-7402 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. guoqing zhu

Verified customer

Submitted Jan 04 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
No blocking step used for 30 minute(s) · Temperature: 25°C
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Pepsin
Sample
Rat Tissue sections (bone)
Specification
bone
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 22 2014

Answer


We would typically suggest using 106 cells in 100 µl diluted antibody. However, the information regarding how ab54741 has come from an Abreview which one of our customers submitted. Unfortunately, they did not provide any details of exactly how the dilution was performed and with how many cells. This will need to be optimised with the particular samples you are using with this antibody for the optimum results. The review can be accessed from the following link:

https://www.abcam.com/rage-antibody-ab54741.html/reviews/12604

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

Answer

Thank you for your enquiry regarding ab54741.

As the datasheet indicates, this antibody has been tested in WB, Flow Cyt, IHC-P. It has not been characterized specifically in IHC-Fr but it is very likely that ab54741 works on frozen sections. Normally we would suggest 1 or 2 hrs incubation with the primary antibody at RT or overnight at 4oC.

Since IHC-Fr is untested application, I could consider offering a discount code for testing. I would advise you to visit the following site where the Terms and Conditions of our testing initiative can be found at: https://www.abcam.com/abtrial

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Answer

Thank you for participating in our recent customer satisfaction survey.

I was in touch with our production lab for ab54741 and they said that the concentration of the antibody is different from batch to batch. Please note that even if you receive a batch that is 0.5 mg/ml instead of 1 mg/ml that we are still sending 100 ug in the vial. The actual mass or amount of the antibody received remains unchanged. You should still see similar labeling, you will just have to account for theconcentration difference when diluting the antibody. If you would prefer to receive a batch at 1 mg/ml in the future, please call our Customer Services department who can check the concentrations of the lots we have in stock and assign the most concentrated one to your order.

If you feel as though the new lot wasn't working at all, even when considering the difference in concentration, then we would be happy to investigate this further. We guarantee that our products will work in the tested species and applications listed on the datasheet and will refund or replace faulty products within 6 months of purchase in accordance with ourAbpromise.

https://www.abcam.com/index.html?pageconfig=abpromise

Please call us (888) 77-ABCAM (22226)or reply to this email and we would be happy to discuss the protocol tested and the results you received with the less concentrated lot. We look forward to helping you and again thank you for your feedback!

Read More

Answer

Thank you for contacting us. It was nice talking to you.

I have discussed the protocol with lab and they have confirmed that they have used microwave antigen retrieval method for 5 minutes with citrate buffer pH 6.0. They have also recommended to use signal enhancer along with normal IHC detection reagents.

For optimization I can recommend trying microwave for 5 minutes and 10 minutes (700 watt, full power).

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

Read More

Question
Answer

Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten.

Ich habe zwei Antikörper gefunden, deren Immunogene eine hohe Homologie mit Kaninchen aufweisen und die schon für WB und IHC-P getestet wurden:

ab65965: 95%

https://www.abcam.com/index.html?datasheet=65965 (or use the following: https://www.abcam.com/index.html?datasheet=65965).

ab86767: 96%

https://www.abcam.com/index.html?datasheet=86767 (or use the following: https://www.abcam.com/index.html?datasheet=86767).



Unserem Wissen nach wurden wederab65965 noch ab86767 bisher im Kaninchen getestet. Falls Sie dies selbst testen möchten, kann ich Ihnen zurzeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert von dem getesteten Antikörper abdecken, und gegen eine erneute Bestellung eines weiterenprimären Antikörpersvon uns verrechnet werden.

Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

1.) Bestätigen Sie mir bitte, dasswelchen Antikörper SieimKaninchen testen möchten.

2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

4.) Testen Sie den Antikörper im Kaninchen.

5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
https://www.abcam.com/abreviews

6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv, und Sie können einen anderenprimären Antikörperzu dem gleichen Preis wieden getestetenbei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode und die Bestellnummer bereit). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
https://www.abcam.com/collaborationdiscount

Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (smooth muscle cells)
Loading amount
50 µg
Specification
smooth muscle cells
Treatment
100 ug/ml glycated LDL
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Blocking step
Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Dr. Laura Toma

Verified customer

Submitted Feb 24 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Recombinant protein (human RAGE cDNA transfected into HEK293 cells)
Loading amount
1 µg
Specification
human RAGE cDNA transfected into HEK293 cells
Gel Running Conditions
Reduced Denaturing (10-20%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Nov 21 2008

1-10 of 11 Abreviews or Q&A

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