Product datasheet
Anti-Glucose Transporter GLUT4 antibody
[mAbcam65267] ab65267
8 Abreviews 2 References 7 Images
Overview
Product name
Anti-Glucose Transporter GLUT4 antibody [mAbcam65267]
Description
Mouse monoclonal [mAbcam65267] to Glucose Transporter GLUT4
Tested applications
IHC-P, WB, ICC/IF, Flow Cyt, IHC-Fr
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Sheep, Rabbit, Goat, Horse, Cow, Pig
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of
Human Glucose Transporter GLUT4.Read Abcam's proprietary immunogen policy(Peptide
available as ab34088.)
Positive control
This antibody gave a positive signal in Mouse Skeletal Muscle, Mouse Heart, Rat Skeletal
Muscle and Rat Heart. This antibody also gave a positive signal against the partial recombinant
GLUT4 protein which has an expected molecular weight of 30 kDa
General notes
We can conjugate this antibody to FITC for you (please see ab150206 for details).
This antibody is available as an azide-free product (see ab176245).
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or 80°C. Avoid freeze / thaw cycle.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: PBS, pH 7.4
Purity
IgG fraction
Clonality
Monoclonal
Clone number
mAbcam65267
Myeloma
Sp2/0
Isotype
IgG2a
Applications
Our Abpromise guarantee covers the use of ab65267 in the following tested applications.
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The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
Notes
IHC-P
Use at an assay dependent concentration.
WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 54 kDa
(predicted molecular weight: 54 kDa).Can be blocked with Human Glucose
Transporter GLUT4 peptide (ab34088).
Abcam recommends using 5% BSA as the blocking agent.
ICC/IF
1/1000.
Flow Cyt
Use 0.5µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use
as an isotype control with this antibody.
IHC-Fr
1/500.
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.
Anti-Glucose Transporter GLUT4 antibody [mAbcam65267] images
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ab65267 stained HeLa cells. The cells were
4% formaldehyde fixed for 10 minutes
at room temperature and then incubated in
1%BSA / 10% normal goat serum / 0.3M
glycine in 0.1% PBS-Tween for 1hour at room
temperature to permeabilise the cells and
block non-specific protein-protein
interactions. The cells were then incubated
with the antibody (ab65267 at 5µg/ml)
overnight at +4°C. The secondary antibody
(pseudo-colored green) was Goat Anti-Mouse
Immunocytochemistry/ Immunofluorescence -
IgG H&L (Alexa Fluor® 488) preadsorbed
Anti-Glucose Transporter GLUT4 antibody
(ab150117) used at a 1/1000 dilution for
[mAbcam65267] (ab65267)
1hour at room temperature. Alexa Fluor® 594
WGA was used to label plasma membranes
(pseudo-colored red) at a 1/200 dilution for
1hour at room temperature. DAPI was used to
stain the cell nuclei (pseudo-colored blue) at a
concentration of 1.43µM for 1hour at room
temperature.
Anti-Glucose Transporter GLUT4 antibody
[mAbcam65267] (ab65267) at 1 µg/ml +
Partial tagged recombinant protein to GLUT4
at 0.1 µg
Secondary
Rabbit polyclonal to Mouse IgG - H&L (HRP)
at 1/3000 dilution
Performed under reducing conditions.
Western blot - Anti-Glucose Transporter GLUT4
Predicted band size : 54 kDa
antibody [mAbcam65267] (ab65267)
Observed band size : 30 kDa
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ab65267 staining glucose transporter GLUT4
in human heart tissue section by
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections). Tissue
underwent fixation in formaldehyde, heat
mediated antigen retrieval in Citrate buffer pH
6.0 and blocking (5 minutes/peroxidase block
then 10 minutes/protein block) for 15 minutes
at 20°C. The primary antibody was diluted,
Immunohistochemistry (Formalin/PFA-fixed
1/2000 and incubated with sample for 45
paraffin-embedded sections) - Glucose
minutes at 20°C. A HRP conjugated goat
Transporter GLUT4 antibody [mAbcam65267]
polyclonal to mouse IgG was used undiluted
(ab65267)
as secondary.
This image is courtesy of an Abreview submitted by
Antibody Solutions Ltd.
All lanes : Anti-Glucose Transporter GLUT4
antibody [mAbcam65267] (ab65267) at 1
µg/ml
Lane 1 : Skeletal Muscle (Mouse) Tissue
Lysate
Lane 2 : Heart (Mouse) Tissue Lysate
Lane 3 : Skeletal Muscle (Rat) Tissue Lysate
Lane 4 : Heart (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Western blot - Glucose Transporter GLUT4
antibody [mAbcam65267] (ab65267)
Secondary
Goat polyclonal to Mouse IgG - H&L - PreAdsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 54 kDa
Observed band size : 54 kDa
Exposure time : 20 minutes
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ab65267 staining glucose transporter GLUT4
in rat heart muscle tissue sections by
Immunohistochemistry (Frozen section).
Tissue was fixed with 4% PFA overnight.
Samples were then blocked with 10% donkey
serum for 1 hour at 24°C followed by
incubation with ab65267 at a 1/500 dilution,
for 24 hours at 4°C. A Donkey anti-rabbit
Alexa Fluoro 568 conjugate was used as
- Glucose Transporter GLUT4 antibody
secondary antibody at a 1/1000 dilution.
[mAbcam65267] (ab65267)
Nuclei were stained using the standard
This image is courtesy of an Abreview submitted by
Ruma Raha-Chowdhury
Hoestch method and are depicted in blue.
ab65267 staining glucose transporter GLUT4
in rat heart muscle tissue sections by
Immunohistochemistry (Frozen section).
Tissue was fixed with 4% PFA overnight.
Samples were then blocked with 10% donkey
serum for 1 hour at 24°C followed by
incubation with ab65267 at a 1/500 dilution,
for 24 hours at 4°C. A Donkey anti-rabbit
Alexa Fluoro 568 conjugate was used as
secondary antibody at a 1/1000 dilution.
Nuclei were stained using the standard
Hoestch method and are depicted in blue.
ab65267 staining cultured mouse striatum
neurons by ICC/IF. The cultured neurons were
fixed with paraformaldehyde, permeabilized
with 0.3% Triton X in 0.1% PBS and blocked
with 10% donkey serum for 30 minutes at
24°C. The cultured neurons were then stained
with ab65267 at 1/1000 in 0.3% TritonX with
0.1% PBS and 10% donkey serum for 4h at
24°C. An Alexa Fluro 568 donkey anti-mouse
Immunocytochemistry/ Immunofluorescence -
polyclonal antibody at 1/1000 was used as
Anti-Glucose Transporter GLUT4 antibody
the secondary antibody.
[mAbcam65267] (ab65267)
This image is courtesy of an Abreview submitted by
Ruma Raha-Chowdhury
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Overlay histogram showing NSO cells stained
with ab65267 (red line). The cells were fixed
with 4% paraformaldehyde (10 min) and then
permeabilized with 0.1% PBS-Tween for 20
min. The cells were then incubated in 1x PBS
/ 10% normal goat serum / 0.3M glycine to
block non-specific protein-protein interactions
followed by the antibody (ab65267,
Flow Cytometry - Anti-Glucose Transporter
0.5µg/1x106 cells) for 30 min at 22ºC. The
GLUT4 antibody [mAbcam65267] (ab65267)
secondary antibody used was DyLight® 488
goat anti-mouse IgG (H+L) (ab96879) at
1/500 dilution for 30 min at 22ºC. Isotype
control antibody (black line) was mouse
IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells)
used under the same conditions. Acquisition
of >5,000 events was performed. This
antibody gave a positive signal in NSO cells
fixed with 80% methanol (5
min)/permeabilized with 0.1% PBS-Tween for
20 min used under the same conditions.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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