Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21171 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
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 |
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IP |
1/30.
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WB | (1) |
1/1000. Predicted molecular weight: 42 kDa.
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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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. |
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ICC/IF |
1/100.
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Flow Cyt |
1/500.
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Notes |
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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
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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
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Database links
- Entrez Gene: 177 Human
- Entrez Gene: 11596 Mouse
- Entrez Gene: 81722 Rat
- Omim: 600214 Human
- SwissProt: Q15109 Human
- SwissProt: Q62151 Mouse
- SwissProt: Q63495 Rat
- Unigene: 534342 Human
see all -
Alternative names
- Advanced glycosylation end product-specific receptor antibody
- Ager antibody
- DAMA 358M23.4 antibody
see all
Images
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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 secondsThe expression profile and molecular mass observed is consistent with what has been described in the literature (PMID:16315007; PMID:18355449; PMID:18245812)
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (9)
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