製品の概要

  • 製品名Anti-RAGE antibody
    RAGE 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to RAGE
  • 特異性By Western blot, this antibody detects two bands in the 45 kDa range representing the RAGE protein pre and post glycosylation in Mouse lung extract. This antibody also detects an ~25 kDa protein that is believed to be proteolytic degradation product. Immunohistochemical staining of RAGE in transgenic Mouse retina results in staining of the retinal pigmented epithelium and photo receptor cell layers.
  • アプリケーション適用あり: ICC/IF, IHC-Fr, WB, Flow Cyt, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to Rat RAGE aa 362-380.
    Sequence:

    WRKRQPR(R/L)EERKAPESQED


    (Peptide available as ab41778)

  • ポジティブ・コントロール
    • Mouse lung extract

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab3611 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF 1/200.
IHC-Fr Use a concentration of 1 - 2 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 45 kDa (predicted molecular weight: 42.6 kDa).Can be blocked with Rat RAGE peptide (ab41778).
Flow Cyt 1/100.
IHC-P 1/10 - 1/100.

ターゲット情報

  • 機能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.
  • 組織特異性Endothelial cells.
  • 配列類似性Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • 細胞内局在Secreted and Cell membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Advanced glycosylation end product-specific receptor antibody
    • Ager antibody
    • MGC2235 antibody
    • RAGE_HUMAN antibody
    • Receptor for advanced glycosylation end products antibody
    see all

Anti-RAGE antibody 画像



  • Predicted band size : 42.6 kDa

    ab3611 at a 2µg/ml concentration staining ~ 45 kDa RAGE in mouse lung lysate by Western blot (ECL).

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Mouse lymph node tissue . To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rabbit polyclonal antibody recognizing RAGE ab3611 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • ab3611 at 1/200 staining mouse BV2 cells by ICC/IF. The cells were formaldehyde fixed, permeabilized and blocked with serum before incubation with the antibody for 1 hour. An Alexa Fluor ® 488 conjugated goat anti-rabbit antibody was used as the secondary. The left hand panel shows unstimulated cells, the right hand panel LPS stimulated cells.

    See Abreview

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Mouse heart tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rabbit polyclonal antibody recognizing RAGE ab3611 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Ab3611 used in IHC (frozen) in transgenic mouse retinas.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Mouse kidney tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rabbit polyclonal antibody recognizing RAGE ab3611 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Anti-RAGE antibody (ab3611) 使用論文

This product has been referenced in:
  • Kim J  et al. Cytoplasmic translocation of high-mobility group box-1 protein is induced by diabetes and high glucose in retinal pericytes. Mol Med Rep 14:3655-61 (2016). Read more (PubMed: 27599553) »
  • Yao X  et al. Mitochondrial ROS Induces Cardiac Inflammation via a Pathway through mtDNA Damage in a Pneumonia-Related Sepsis Model. PLoS One 10:e0139416 (2015). IHC ; Rat . Read more (PubMed: 26448624) »

See all 23 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Lung endothelial cells)
Permeabilization Yes - 0.5% Triton in PBS
Specification Lung endothelial cells
Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Acetone:Methanol
Username

Jose Vazquez-Medina

Verified customer

投稿 Nov 14 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Whole lung tissue lysate)
Gel Running Conditions Reduced Denaturing (4-12% BT gel)
Loading amount 25 µg
Specification Whole lung tissue lysate
Blocking step Odyssey Blocking Buffer (PBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Aug 25 2016

Application Western blot
Loading amount 20 µg
Gel Running Conditions Non-reduced Denaturing (12.5% SDS-PAGE)
Sample Mouse Tissue lysate - whole (mouse lung)
Specification mouse lung
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Sep 12 2014

We have not stained dot blots with this antibody but it stains western blots effectively (reduced/denatured samples). I think a western blot would be more convincing data, for example the customer review at this link.

http://www.abcam.com/RAGE...

Read More
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Antigen retrieval step None
Sample Human Tissue sections (skin)
Specification skin
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Jun 11 2014

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (RAW 264.7)
Specification RAW 264.7
Permeabilization Yes - 0.1% Triton X-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Oct 18 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunoprecipitation
Sample Mouse Cell lysate - whole cell (RAW 264.7)
Total protein in input 500 µg
Treatment In log phase growth in 10% FCS
Immuno-precipitation step Other - PureProteome Protein G Magnetic Beads
Specification RAW 264.7
Username

Abcam user community

Verified customer

投稿 Oct 17 2013

Thank you for contacting us. I have recieved the following expnation about observed bands from laboratory.

The two bands in the 45kD range are expected because RAGE is a glycosylated protein. Based on the expected MW of RAGE, it is likely that...

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Thank you for your email.

This antibody is raised against immunogen aa 362-380 of human RAGE. The immunogen sequence corresponds to Isoform 1 only, though there are 4 know isoforms of this protein as given in Swissprot database (http://www.abc...

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Thank you for this additional information. Your protocol seems sound but you may want to try increasing your Triton to 0.25% while also increasing the blocking to 10% for one hour. This may give you a better signal.
We do guarantee our products for...

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