Overview

  • Product name
    Anti-Glucose Transporter GLUT4 antibody
    See all Glucose Transporter GLUT4 primary antibodies
  • Description
    Rabbit polyclonal to Glucose Transporter GLUT4
  • Host species
    Rabbit
  • Specificity
    GLUT4.
  • Tested applications
    Suitable for: ICC/IF, WB, IP, Immunomicroscopy, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Pig, Armenian hamster
  • Immunogen

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

  • Positive control
    • IHC-P: FFPE human heart tissue sections.

Properties

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.

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

  • Function
    Insulin-regulated facilitative glucose transporter.
  • Tissue specificity
    Skeletal and cardiac muscles; brown and white fat.
  • Involvement in disease
    Diabetes mellitus, non-insulin-dependent
  • Sequence similarities
    Belongs to the major facilitator superfamily. Sugar transporter (TC 2.A.1.1) family. Glucose transporter subfamily.
  • Post-translational
    modifications
    Sumoylated.
  • Cellular localization
    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
  • Database links
  • Alternative names
    • 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

Images

  • 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

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) »
See all 96 Publications for this product

Customer reviews and Q&As

1-10 of 48 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Submitted 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

Submitted 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

Submitted 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
Abcam guarantees this product to work in the species/application used in this Abreview.
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

Submitted Jun 10 2013

Answer

Hello and thank you for sending your image. We do highly recommend boiling your samples with this antibody, and this may reduce background. However, this target is phosphorylated and glycosylated which is another reason you may see two strong bands. If you do not wish to boil the samples you may want to use the blocking peptide ab115831 to confirm that these bands are GLUT4 specific. Please let me know if you have any further questions or concerns.

Read More

Answer

Thank you for your new contact. My colleague is out of office today and she has asked me to look after her customers.

Currently, we do not have a different Western blot image available.The predicted molecular weight of GLUT4 is54.8 kDa, however due to post-translational modification (N-glycosylation) the mature protein may have a signal at the higher size.

If you need any further assistance in the future, please do not hesitate to contact me.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Mesenchymal stem cells)
Specification
Mesenchymal stem cells
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C

Mr. Steven Johnstone

Verified customer

Submitted Sep 11 2012

Answer

Thank you for contacting us.

All the images from WB assay in the datasheet belong to Abreviews from other users. I am afraid we do not have access to the complete images.

As new images or references are brought to our attention, these will be updated on the datasheet.

This antibody is guaranteed to work in with WB. If you had any problems using it in such application, we would them send you a replacement or reimbursement as guaranteed by the Abpromise guarantee.

I am very sorry I couldn’t help you further. Please do not hesitate to contact us if you need any other advice or information.

Read More

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge blocking buffer to use in your western blots with the order number xxxxxx. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

Read More

1-10 of 48 Abreviews or Q&A

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