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RecombinantRabMAb

Recombinant Anti-RAGE antibody [EPR21171] (ab216329)

Reviews (1) Submit a question References (9)
Western blot - Anti-RAGE antibody [EPR21171] (ab216329)
  • Immunohistochemistry (Frozen sections) - Anti-RAGE antibody [EPR21171] (ab216329)
  • Western blot - Anti-RAGE antibody [EPR21171] (ab216329)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [EPR21171] (ab216329)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [EPR21171] (ab216329)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [EPR21171] (ab216329)
  • Immunohistochemistry (Frozen sections) - Anti-RAGE antibody [EPR21171] (ab216329)
  • Immunocytochemistry/ Immunofluorescence - Anti-RAGE antibody [EPR21171] (ab216329)
  • Flow Cytometry - Anti-RAGE antibody [EPR21171] (ab216329)
  • Immunoprecipitation - Anti-RAGE antibody [EPR21171] (ab216329)
  • Anti-RAGE antibody [EPR21171] (ab216329)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21171] to RAGE
  • Suitable for: IP, WB, IHC-P, IHC-Fr, ICC/IF, Flow Cyt
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free PE

Overview

  • Product name

    Anti-RAGE antibody [EPR21171]
    See all RAGE primary antibodies

  • Description

    Rabbit monoclonal [EPR21171] to RAGE

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WB, IHC-P, IHC-Fr, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal lung lysate; Mouse and rat lung lysates. IHC-P: Human, mouse and rat lung tissues. IHC-Fr: Mouse and rat lung tissues. ICC/IF: HEK-293T cells. Flow Cyt: HEK-293T cells. IP: Mouse lung lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab216329 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
IP
1/30.
WB (1)
1/1000. Predicted molecular weight: 42 kDa.
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr
1/500.

Perform heat mediated antigen retrieval by using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) before commencing with IHC staining protocol.
ICC/IF
1/100.
Flow Cyt
1/500.
Notes
IP
1/30.
WB
1/1000. Predicted molecular weight: 42 kDa.
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr
1/500.

Perform heat mediated antigen retrieval by using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20) before commencing with IHC staining protocol.
ICC/IF
1/100.
Flow Cyt
1/500.

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

    see all

  • 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

  • Western blot - Anti-RAGE antibody [EPR21171] (ab216329)
    Western blot - Anti-RAGE antibody [EPR21171] (ab216329)
    All lanes : Anti-RAGE antibody [EPR21171] (ab216329) at 1/2000 dilution

    Lane 1 : Mouse lung lysates with 5% NFDM/TBST
    Lane 2 : Mouse brain lysates with 5% NFDM/TBST
    Lane 3 : Mouse kidney lysates with 5% NFDM/TBST
    Lane 4 : Mouse heart lysates with 5% NFDM/TBST
    Lane 5 : Mouse liver lysates with 5% NFDM/TBST
    Lane 6 : Mouse spleen lysates with 5% NFDM/TBST
    Lane 7 : Rat lung lysates with 5% NFDM/TBST
    Lane 8 : Rat brain lysates with 5% NFDM/TBST
    Lane 9 : Rat kidney lysates with 5% NFDM/TBST
    Lane 10 : Rat heart lysates with 5% NFDM/TBST
    Lane 11 : Rat spleen lysates with 5% NFDM/TBST

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 42 kDa
    Observed band size: 43 kDa why is the actual band size different from the predicted?


    Exposure time: 3 seconds


    The expression profile and molecular mass observed is consistent with what has been described in the literature (PMID:16315007; PMID:18355449; PMID:18245812)

  • Immunohistochemistry (Frozen sections) - Anti-RAGE antibody [EPR21171] (ab216329)
    Immunohistochemistry (Frozen sections) - Anti-RAGE antibody [EPR21171] (ab216329)

    Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen rat lung tissue labeling RAGE with ab216329 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive membrane staining on alveolar epithelial cells, negative on the bronchial epithelial cells on rat lung tissue section is observed (PMID: 15173891).

    The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Western blot - Anti-RAGE antibody [EPR21171] (ab216329)
    Western blot - Anti-RAGE antibody [EPR21171] (ab216329)
    All lanes : Anti-RAGE antibody [EPR21171] (ab216329) at 1/1000 dilution

    Lane 1 : Mouse lung lysate
    Lane 2 : Rat lung lysate
    Lane 3 : Human fetal lung lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lane 3 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Developed using the ECL technique.

    Predicted band size: 42 kDa
    Observed band size: 45,55 kDa why is the actual band size different from the predicted?



    Exposure times: Lane 1: 5 seconds; Lane 2: 10 seconds; Lane 3: 3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

    The expression profile and molecular mass observed is consistent with what has been described in the literature (PMID:16315007; PMID:18355449; PMID:18245812). Full-length RAGE is not detected in rat and human lysates.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [EPR21171] (ab216329)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [EPR21171] (ab216329)

    Immunohistochemical analysis of paraffin-embedded human lung tissue labeling RAGE with ab216329 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on epithelial cells of human lung (PMID: 19592063; PMID: 26472810) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [EPR21171] (ab216329)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [EPR21171] (ab216329)

    Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling RAGE with ab216329 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Mainly membranous staining on epithelial cells of mouse lung (PMID: 19592063; PMID: 26472810) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [EPR21171] (ab216329)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAGE antibody [EPR21171] (ab216329)

    Immunohistochemical analysis of paraffin-embedded rat lung tissue labeling RAGE with ab216329 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Mainly membranous staining on epithelial cells of rat lung (PMID: 19592063; PMID: 26472810) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Frozen sections) - Anti-RAGE antibody [EPR21171] (ab216329)
    Immunohistochemistry (Frozen sections) - Anti-RAGE antibody [EPR21171] (ab216329)

    Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse lung tissue labeling RAGE with ab216329 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Positive membrane staining on alveolar epithelial cells, negative on the bronchial epithelial cells on mouse lung tissue section is observed (PMID: 15173891).

    The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-RAGE antibody [EPR21171] (ab216329)
    Immunocytochemistry/ Immunofluorescence - Anti-RAGE antibody [EPR21171] (ab216329)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with Myc-tagged RAGE expression vector labeling RAGE with ab216329 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing positive staining in HEK-293T cells transfected with Myc-tagged RAGE expression vector. 

    The nuclear counter stain is DAPI (blue). Myc-Tag is detected with Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 647 Conjugate) (red) at 1/1000 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    Negative control: Myc-transfected HEK-293T cells.

  • Flow Cytometry - Anti-RAGE antibody [EPR21171] (ab216329)
    Flow Cytometry - Anti-RAGE antibody [EPR21171] (ab216329)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with Myc-tagged RAGE expression vector labeling RAGE with ab216329 at 1/500 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    Fresh cells were surface-stained with ab172730 and ab216329 respectively. Then fixed with 2% PFA for 15min and intracellular stained with anti-Myc tag antibody (Y axis). Only Myc+ population give positive signal.

  • Immunoprecipitation - Anti-RAGE antibody [EPR21171] (ab216329)
    Immunoprecipitation - Anti-RAGE antibody [EPR21171] (ab216329)

    RAGE was immunoprecipitated from 0.35 mg of mouse lung lysate with ab216329 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216329 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

    Lane 1: Mouse lung lysate 10 µg (Input). 
    Lane 2: ab216329 IP in mouse lung lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216329 in mouse lung lysate.

     

    Exposure time: 10 seconds.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • Anti-RAGE antibody [EPR21171] (ab216329)
    Anti-RAGE antibody [EPR21171] (ab216329)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (9)

Publishing research using ab216329? Please let us know so that we can cite the reference in this datasheet.

ab216329 has been referenced in 9 publications.

  • He C  et al. The role of irreversible electroporation in promoting M1 macrophage polarization via regulating the HMGB1-RAGE-MAPK axis in pancreatic cancer. Oncoimmunology 10:1897295 (2021). PubMed: 33763295
  • Chen Q  et al. Rho-kinase inhibitor hydroxyfasudil protects against HIV-1 Tat-induced dysfunction of tight junction and neprilysin/Aß transfer receptor expression in mouse brain microvessels. Mol Cell Biochem 476:2159-2170 (2021). PubMed: 33548010
  • Liu M  et al. Progranulin Protects Against Airway Remodeling Through the Modulation of Autophagy via HMGB1 Suppression in House Dust Mite-Induced Chronic Asthma. J Inflamm Res 14:3891-3904 (2021). PubMed: 34408470
  • Haraguchi R  et al. New Insights into the Pathogenesis of Diabetic Nephropathy: Proximal Renal Tubules Are Primary Target of Oxidative Stress in Diabetic Kidney. Acta Histochem Cytochem 53:21-31 (2020). PubMed: 32410750
  • Liang Y  et al. Inhibition of Caspase-1 Ameliorates Ischemia-Associated Blood-Brain Barrier Dysfunction and Integrity by Suppressing Pyroptosis Activation. Front Cell Neurosci 14:540669 (2020). PubMed: 33584203
  • Tsoporis JN  et al. Circulating Ligands of the Receptor for Advanced Glycation End Products and the Soluble Form of the Receptor Modulate Cardiovascular Cell Apoptosis in Diabetes. Molecules 25:N/A (2020). PubMed: 33182705
  • Sun Z  et al. RAGE/galectin-3 yields intraplaque calcification transformation via sortilin. Acta Diabetol N/A:N/A (2019). PubMed: 30603868
  • Shi M  et al. The RAGE/STAT5/autophagy axis regulates senescence in mesangial cells. Cell Signal 62:109334 (2019). PubMed: 31158467
  • Zhao Y  et al. High glucose-induced complement component 3 up-regulation via RAGE-p38MAPK-NF-?B signalling in astrocytes: In vivo and in vitro studies. J Cell Mol Med 22:6087-6098 (2018). PubMed: 30246940

Customer reviews and Q&As

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Western blot abreview for Anti-RAGE antibody [EPR21171]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (MDA-MB-231 cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin gel)
Loading amount
40 µg
Specification
MDA-MB-231 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Apr 26 2022

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