RAGE antibody [EPR21171] (ab216329)

Product name

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

Rabbit monoclonal [EPR21171] to RAGE
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-Fr	Mouse Rat
IHC-P	Mouse Rat Human
IP	Mouse
WB	Mouse Rat Human

See all applications and species data

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.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR21171
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human RAGE protein (Fc Chimera Active) (ab219875)
Related Products
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-Fr	Mouse Rat
IHC-P	Mouse Rat Human
IP	Mouse
WB	Mouse Rat Human

Application	Abreviews	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.

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.

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.
Target information above from: UniProt accession Q15109 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
- Unigene: 3383 Mouse
- Unigene: 9829 Rat
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

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)

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)

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)

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)

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)

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)

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)

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 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)

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)

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

Datasheet

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

ab216329 has been referenced in 4 publications.

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
Shi M et al. The RAGE/STAT5/autophagy axis regulates senescence in mesangial cells. Cell Signal 62:109334 (2019). PubMed: 31158467
Sun Z et al. RAGE/galectin-3 yields intraplaque calcification transformation via sortilin. Acta Diabetol N/A:N/A (2019). PubMed: 30603868
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

Recombinant Anti-RAGE antibody [EPR21171] (ab216329)

Key features and details

Overview

Product name

Description

Host species

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Certificate of Compliance

References (4)

Customer reviews and Q&As