Anti-Glucose Transporter GLUT4 antibody (ab654)
- Datasheet
- References (96)
- Protocols
Overview
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Product nameAnti-Glucose Transporter GLUT4 antibody
See all Glucose Transporter GLUT4 primary antibodies -
DescriptionRabbit polyclonal to Glucose Transporter GLUT4
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Host speciesRabbit
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SpecificityGLUT4.
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Tested applicationsSuitable for: ICC/IF, WB, IP, Immunomicroscopy, IHC-Fr, IHC-Pmore details
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Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Pig, Armenian hamster
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Immunogen
Synthetic peptide corresponding to Rat Glucose Transporter GLUT4 aa 495-509 (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available asab115831) -
Positive control
- IHC-P: FFPE human heart tissue sections.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Storage bufferPreservative: 0.01% Sodium azide
Constituent: Whole serum -
PurityWhole antiserum
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Related Products
Applications
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.
| Application | Abreviews | Notes |
|---|---|---|
| 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. |
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| 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. |
Target
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FunctionInsulin-regulated facilitative glucose transporter.
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Tissue specificitySkeletal and cardiac muscles; brown and white fat.
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Involvement in diseaseDiabetes mellitus, non-insulin-dependent
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Sequence similaritiesBelongs to the major facilitator superfamily. Sugar transporter (TC 2.A.1.1) family. Glucose transporter subfamily.
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Post-translational
modificationsSumoylated. -
Cellular localizationCell 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
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Database links
- Entrez Gene: 6517 Human
- Entrez Gene: 20528 Mouse
- Entrez Gene: 25139 Rat
- Omim: 138190 Human
- SwissProt: P14672 Human
- SwissProt: P14142 Mouse
- SwissProt: P19357 Rat
- Unigene: 380691 Human
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Alternative names
- insulin-responsive antibody
- Glucose transporter GLUT 4 antibody
- Glucose transporter type 4 antibody
see all
Images
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Western blot - Anti-Glucose Transporter GLUT4 antibody (ab654)Image courtesy of an anonymous AbreviewAnti-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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT4 antibody (ab654)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
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Immunocytochemistry/ Immunofluorescence - Anti-Glucose Transporter GLUT4 antibody (ab654)This image is courtesy of Steven Johnstoneab654 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).
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Immunohistochemistry (Frozen sections) - Anti-Glucose Transporter GLUT4 antibody (ab654)ab654 at a 1/300 dilution, staining human GLUT4 in fetal heart tissue by Immunohistochemistry, frozen tissue. -
Immunocytochemistry/ Immunofluorescence - Anti-Glucose Transporter GLUT4 antibody (ab654)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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT4 antibody (ab654)This image is courtesy of an abreview submitted by Brett McKinnonab654 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.
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Immunohistochemistry (Frozen sections) - Anti-Glucose Transporter GLUT4 antibody (ab654)This image is courtesy of an anonymous 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.
Protocols
References
This product has been referenced in:
- Zhang M et al. Semen Cassiae Extract Improves Glucose Metabolism by Promoting GlUT4 Translocation in the Skeletal Muscle of Diabetic Rats. Front Pharmacol 9:235 (2018). Read more (PubMed: 29670524) »
- Tetri LH et al. Sex-Specific Skeletal Muscle Fatigability and Decreased Mitochondrial Oxidative Capacity in Adult Rats Exposed to Postnatal Hyperoxia. Front Physiol 9:326 (2018). WB ; Rat . Read more (PubMed: 29651255) »
