Anti-Glucose Transporter GLUT1 antibody (ab32551)

Overview

  • Product name
    Anti-Glucose Transporter GLUT1 antibody
    See all Glucose Transporter GLUT1 primary antibodies
  • Description
    Rabbit polyclonal to Glucose Transporter GLUT1
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Rabbit
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Glucose Transporter GLUT1.

    (Peptide available as ab33006.)

  • Positive control
    • ab32551 has given positive results in the following Human Whole Cell Lysates: HeLa Jurkat A431 HEK 293 Mouse Whole Cell Lysates NIH 3T3 MEF1 Mouse Tissue Lysates Liver Kidney Testis Spinal cord Ovary Rat Tissue Lysate Liver

Properties

Applications

Our Abpromise guarantee covers the use of ab32551 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 1/200.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 55 kDa).
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use a concentration of 5 µg/ml.

Target

  • Function
    Facilitative glucose transporter. This isoform may be responsible for constitutive or basal glucose uptake. Has a very broad substrate specificity; can transport a wide range of aldoses including both pentoses and hexoses.
  • Tissue specificity
    Expressed at variable levels in many human tissues.
  • Involvement in disease
    Defects in SLC2A1 are the cause of glucose transporter type 1 deficiency syndrome (GLUT1DS) [MIM:606777]; also known as blood-brain barrier glucose transport defect. This disease causes a defect in glucose transport across the blood-brain barrier. It is characterized by infantile seizures, delayed development, and acquired microcephaly.
    Defects in SLC2A1 are the cause of dystonia type 18 (DYT18) [MIM:612126]. DYT18 is an exercise-induced paroxysmal dystonia/dyskinesia. Dystonia is defined by the presence of sustained involuntary muscle contraction, often leading to abnormal postures. DYT18 is characterized by attacks of involuntary movements triggered by certain stimuli such as sudden movement or prolonged exercise. In some patients involuntary exertion-induced dystonic, choreoathetotic, and ballistic movements may be associated with macrocytic hemolytic anemia.
  • Sequence similarities
    Belongs to the major facilitator superfamily. Sugar transporter (TC 2.A.1.1) family. Glucose transporter subfamily.
  • Post-translational
    modifications
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization
    Cell membrane. Melanosome. Localizes primarily at the cell surface (By similarity). Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
  • Alternative names
    • Choreoathetosis/spasticity episodic (paroxysmal choreoathetosis/spasticity) antibody
    • CSE antibody
    • DYT17 antibody
    • DYT18 antibody
    • DYT9 antibody
    • EIG12 antibody
    • erythrocyte/brain antibody
    • Erythrocyte/hepatoma glucose transporter antibody
    • facilitated glucose transporter member 1 antibody
    • Glucose transporter 1 antibody
    • Glucose transporter type 1 antibody
    • Glucose transporter type 1, erythrocyte/brain antibody
    • GLUT antibody
    • GLUT-1 antibody
    • GLUT1 antibody
    • GLUT1DS antibody
    • GLUTB antibody
    • GT1 antibody
    • GTG1 antibody
    • Gtg3 antibody
    • GTR1_HUMAN antibody
    • HepG2 glucose transporter antibody
    • HTLVR antibody
    • Human T cell leukemia virus (I and II) receptor antibody
    • MGC141895 antibody
    • MGC141896 antibody
    • PED antibody
    • RATGTG1 antibody
    • Receptor for HTLV 1 and HTLV 2 antibody
    • SLC2A1 antibody
    • Solute carrier family 2 (facilitated glucose transporter), member 1 antibody
    • Solute carrier family 2 antibody
    • Solute carrier family 2, facilitated glucose transporter member 1 antibody
    see all

Images

  • All lanes : Anti-Glucose Transporter GLUT1 antibody (ab32551) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse) Whole Cell Lysate
    Lane 3 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 55 kDa
    Observed band size : 51 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 35 kDa,37 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab32551 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

    Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

  • IHC image of Glucose Transporter GLUT1 staining in human kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab32551, 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.
  • ICC/IF image of ab32551 stained HepG2 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 (ab32551, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
  • Glucose Transporter GLUT1 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to Glucose Transporter GLUT1 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32551.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 51kDa; non specific band - 37kDa: We are unsure as to the identity of this extra band. Glucose Transporter GLUT1
  • All lanes : Anti-Glucose Transporter GLUT1 antibody (ab32551) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)
    Lane 4 : HEK293 whole cell lysate (ab7902)

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

    Performed under reducing conditions.

    Predicted band size : 55 kDa
    Observed band size : 50 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 90 kDa. We are unsure as to the identity of these extra bands.
  • All lanes : Anti-Glucose Transporter GLUT1 antibody (ab32551) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Liver (Mouse) Tissue Lysate - normal tissue
    Lane 4 : Kidney (Mouse) Tissue Lysate
    Lane 5 : Testis (Mouse) Tissue Lysate - normal tissue
    Lane 6 : Spinal Cord (Mouse) Tissue Lysate
    Lane 7 : Ovary (Mouse) Tissue Lysate - normal tissue
    Lane 8 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 55 kDa
    Observed band size : 51 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 34 kDa. We are unsure as to the identity of these extra bands.

References

This product has been referenced in:
  • Calado SM  et al. GLUT1 activity contributes to the impairment of PEDF secretion by the RPE. Mol Vis 22:761-70 (2016). WB ; Human . Read more (PubMed: 27440994) »
  • Liemburg-Apers DC  et al. Acute stimulation of glucose influx upon mitoenergetic dysfunction requires LKB1, AMPK, Sirt2 and mTOR-RAPTOR. J Cell Sci 129:4411-4423 (2016). Read more (PubMed: 27793977) »

See all 17 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Mouse Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
20 µg
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Jan 11 2016

Thank you for contacting us.

The immunogen used to generate ab32551 is intracellular. For GLUT1 ab14683, the immunogen corresponds to an extracellular portion, but this antibody hasn't been tested in FC. For our two GLUT1 antibodies tested i...

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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 cau...

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Thanks for getting back to me and letting me know about the results.

I'm sorry to hear that the glycerol actually made the results worse! I've heard some good feedback for that technique and was hoping that it would be helpful. I'm also sorry...

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Abcam has not validated the combination of species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (Peripheral blood leukocytes)
Specification
Peripheral blood leukocytes
Preparation
Cell harvesting/tissue preparation method: Red cell lysis using Ammonium chloride. Wash with FACS flow BSA
Sample buffer: FACS flow
Permeabilization
No
Gating Strategy
cells were gated based on CD19 and CD5 expression
Username

Dr. Abraham Varghese

Verified customer

Submitted Jan 21 2011

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (cerebellum)
Specification
cerebellum
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Oct 23 2008

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (cerebellum saggital section)
Specification
cerebellum saggital section
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Oct 13 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (vascular smooth muscle cells, macrophages)
Loading amount
4 µg
Specification
vascular smooth muscle cells, macrophages
Gel Running Conditions
Reduced Denaturing (gradient 4-12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
Username

Abcam user community

Verified customer

Submitted Jun 12 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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