製品の概要

  • 製品名

    Anti-Glucose Transporter GLUT4 antibody
    Glucose Transporter GLUT4 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to Glucose Transporter GLUT4
  • 由来種

    Rabbit
  • 特異性

    GLUT4.
  • アプリケーション

    適用あり: ICC/IF, WB, IP, Immunomicroscopy, IHC-Fr, IHC-Pmore details
  • 種交差性

    交差種: Mouse, Rat, Human
    交差が予測される動物種: Pig, Armenian hamster
  • 免疫原

    Synthetic peptide corresponding to Rat Glucose Transporter GLUT4 aa 495-509 (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab115831)

  • ポジティブ・コントロール

    • IHC-P: FFPE human heart tissue sections.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab654 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 1/1000.
WB 1/2000 - 1/2500. Detects a band of approximately 45 kDa (predicted molecular weight: 54.8 kDa).Can be blocked with Rat Glucose Transporter GLUT4 peptide (ab115831).

The samples MUST NOT be boiled before running the gel as highly hydrophobic membrane proteins, like glucose transporters, aggregate severely at high temperatures due to the hydrophobic effect, which increases as a direct function of temperature. If the samples are boiled, much of the transporter will aggregate so severely it doesn't enter the running gel and higher order oligomers will be observed.

It is recomended to use 3% milk block.

IP Use at an assay dependent concentration. PubMed: 18234908
Immunomicroscopy Use at an assay dependent concentration. Can also can be used for Immunogold EM (purified IgG). See references for specific protocols.
IHC-Fr 1/500.
IHC-P Use at an assay dependent concentration.

ターゲット情報

  • 機能

    Insulin-regulated facilitative glucose transporter.
  • 組織特異性

    Skeletal and cardiac muscles; brown and white fat.
  • 関連疾患

    Diabetes mellitus, non-insulin-dependent
  • 配列類似性

    Belongs to the major facilitator superfamily. Sugar transporter (TC 2.A.1.1) family. Glucose transporter subfamily.
  • 翻訳後修飾

    Sumoylated.
  • 細胞内局在

    Cell membrane. Endomembrane system. Cytoplasm, perinuclear region. Localizes primarily to the perinuclear region, undergoing continued recycling to the plasma membrane where it is rapidly reinternalized. The dileucine internalization motif is critical for intracellular sequestration.
  • Information by UniProt
  • 参照データベース

  • 別名

    • insulin-responsive antibody
    • Glucose transporter GLUT 4 antibody
    • Glucose transporter type 4 antibody
    • Glucose transporter type 4 insulin responsive antibody
    • GLUT 4 antibody
    • GLUT-4 antibody
    • GLUT4 antibody
    • GTR4_HUMAN antibody
    • Insulin responsive glucose transporter type 4 antibody
    • kug antibody
    • SLC 2A4 antibody
    • SLC2A4 antibody
    • solute carrier family 2 (facilitated glucose transporter) member 4 antibody
    • Solute carrier family 2 member 4 antibody
    • Solute carrier family 2, facilitated glucose transporter member 4 antibody
    see all

画像

  • Anti-Glucose Transporter GLUT4 antibody (ab654) at 1/1000 dilution (Diluted with TBST and Incubated for 16 hours at 4°C) + Mouse Heart at 30 µg with Milk, 1 hour at 25°C at 3 %

    Secondary
    Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) HRP at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 54.8 kDa
    Additional bands at: 45 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 5 seconds

    See Abreview

  • IHC image of Glucose Transporter GLUT4 staining in human heart formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab654, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • ab654 staining GLUT4 in human mesenchymal stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 30 minutes at 22°C. Samples were incubated with primary antibody (1/100 in 10% Horse serum) for 1 hour at 22°C. A FITC-conjugated rabbit polyclonal was used as the secondary antibody (1/100).

    See Abreview

  • ab654 at a 1/300 dilution, staining human GLUT4 in fetal heart tissue by Immunohistochemistry, frozen tissue.

    See Abreview

  • ICC/IF image of ab654 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab654, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • ab654 staining GLUT4 in human endometriosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 3% BSA for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/50 in 3% BSA in TBS) for 16 hours at 4°C. An undiluted Biotin-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • ab654 at 1/500 dilution staining GLUT4 in mouse muscle by immunohistochemistry (frozen sections). Sections were methanol fixed prior to blocking in 5% goat serum for 1 hour at 25°C and then incubated with ab654 for 1 hour at 25°C. A goat polyclonal to rabbit IgG, diluted 1/500, was used as the secondary antibody.

    See Abreview

参考文献

This product has been referenced in:

  • Vassilakos G  et al. Deletion of muscle IGF-I transiently impairs growth and progressively disrupts glucose homeostasis in male mice. FASEB J 33:181-194 (2019). Read more (PubMed: 29932867) »
  • Chen FF  et al. Differences between porcine longissimus thoracis and semitendinosus intramuscular fat content and the regulation of their preadipocytes during adipogenic differentiation. Meat Sci 147:116-126 (2019). Read more (PubMed: 30219363) »
See all 113 Publications for this product

レビューと Q&A

1-10 of 53 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (stomach)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
stomach
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

投稿 Dec 06 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Ileum)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
Ileum
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

投稿 Dec 06 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast cancer)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
Breast cancer
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

投稿 Dec 06 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Artery)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
Artery
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

投稿 Dec 06 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
10mM Sodium Citrate pH6

Abcam user community

Verified customer

投稿 Dec 06 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Placental Explant)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer, pH 6.0
Permeabilization
No
Specification
Placental Explant
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Paraformaldehyde

Jeremiah Gaudet

Verified customer

投稿 Aug 01 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse brain, heart, liver, muscle, and retina)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
30 µg
Specification
Mouse brain, heart, liver, muscle, and retina
Blocking step
Licor Odyssey Blocking BUffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

投稿 Aug 23 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (heart)
Gel Running Conditions
Reduced Denaturing (gel 4-15% SDS)
Loading amount
30 µg
Specification
heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

投稿 Aug 04 2015

Answer

I would recommend looking at the human protein atlas which shows in which tissues the protein is expressed:

http://www.proteinatlas.org/ENSG00000181856/tissue

From this, it would seem that the following tissues could be good negative controls:

Liver, gallbladder, pancreas, oral mucosa, esophagus, duodenum, small intestine, appendix, rectum, epididymis, prostate, seminal vesicle, breast, vagina, cervix, uterus, fallopian tube, placenta, skin, bone marrow, lymph node, tonsil, spleen, thyroid gland, and nasopharynx.

There therefore seems to be a large choice of negative control tissues but I would also recommend cross-checking these with the literature to see if they are true negative controls.

We also sell the blocking peptide for ab654 which you could use as a second negative control: Rat Glucose Transporter GLUT4 peptide (ab115831)

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: 3% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Sample
Human Tissue sections (Endometriosis)
Specification
Endometriosis
Permeabilization
No
Fixative
Paraformaldehyde

Dr. Brett Mckinnon

Verified customer

投稿 Jun 10 2013

1-10 of 53 Abreviews or Q&A

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