Name: Anti-Glucose Transporter GLUT1 antibody [EPR3915]
SKU: ab115730
Rating: 4 (19 reviews)

Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) is a rabbit monoclonal antibody detecting Glucose Transporter GLUT1 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse.

- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

- Over 320 publications

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HRP Anti-Glucose Transporter GLUT1 antibody [EPR3915]
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Anti-Glucose Transporter GLUT1 antibody [EPR3915] - BSA and Azide free
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Alexa Fluor® 405 Anti-Glucose Transporter GLUT1 antibody [EPR3915]
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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (AB115730), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested
Mouse	Not recommended	Tested	Expected	Expected
Rat	Not recommended	Predicted	Predicted	Predicted

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250 - 1/500	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100000	Notes We would not recommend boiling due to the possible irreversible aggregation of glycose transporters. If samples are boiled it can prevent some of the protein from entering the gel or produce multimers which are often mistaken for background. Samples should be solubilized in standard SDS Laemmli buffer and maintained at room temperature before loading.
Species Mouse	Dilution info 1/100000	Notes We would not recommend boiling due to the possible irreversible aggregation of glycose transporters. If samples are boiled it can prevent some of the protein from entering the gel or produce multimers which are often mistaken for background. Samples should be solubilized in standard SDS Laemmli buffer and maintained at room temperature before loading.

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1 µg/mL	Notes This product gave a positive signal in A549 (SLC2A1 knockout A549 cells used as a negative control) fixed with 100% methanol (5 min).

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/40	Notes For unpurified, use 1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Associated Products

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1 product for Alternative Version

Target data

See full target information SLC2A1

Function

Facilitative glucose transporter, which is responsible for constitutive or basal glucose uptake (PubMed:10227690, PubMed:10954735, PubMed:18245775, PubMed:19449892, PubMed:25982116, PubMed:27078104, PubMed:32860739). Has a very broad substrate specificity; can transport a wide range of aldoses including both pentoses and hexoses (PubMed:18245775, PubMed:19449892). Most important energy carrier of the brain: present at the blood-brain barrier and assures the energy-independent, facilitative transport of glucose into the brain (PubMed:10227690). In association with BSG and NXNL1, promotes retinal cone survival by increasing glucose uptake into photoreceptors (By similarity). Required for mesendoderm differentiation (By similarity).

Alternative names

GLUT1, SLC2A1, HepG2 glucose transporter, GLUT-1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR3915
Purification technique: Affinity purification Protein A
Specificity: We recommend not to boil the samples after lysis to get desired WB bands.
Dissociation constant: 7.7 x 10^-12 M
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P and WB.

Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) was first used in a scientific publication in 2012 and has been cited over 320 times in peer reviewed journals. It's performance in Western Blot in human and mouse samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

The specificity of Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) has been confirmed by Western Blot testing in Glucose Transporter GLUT1 knockout HepG2 cells (Human SLC2A1 (Glucose Transporter GLUT1) knockout Hep G2 cell line ab280797).

Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) has 18 independent reviews from customers.

Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) specifically detects Glucose Transporter GLUT1 (UniProt ID: P11166; Molecular weight: 55kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).

Conjugation-ready, carrier free format available for antibody clone EPR3915 - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free ab196357.

Antibody clone EPR3915 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 647, HRP, Alexa Fluor^® 488, Alexa Fluor^® 594, PE, Alexa Fluor^® 405, APC, FITC (Alexa Fluor® 647 Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab195020, HRP Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab195021, Alexa Fluor® 488 Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab195359, Alexa Fluor® 594 Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab206360, PE Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab209449, Alexa Fluor® 405 Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab210438, APC Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab316298, FITC Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab322306).

GLUT1 significantly impacts inflammation by facilitating glucose uptake in immune cells, which is essential for their activation and function. GLUT1 activity can lead to chronic inflammation and tissue damage. GLUT1 is a key biomarker in IBD and immunometabolism. GLUT1 is a signature in Met-flow analysis. Antibody is top cited with over 428 publications.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The Glucose Transporter GLUT1 also known as SLC2A1 is an important protein responsible for the transport of glucose across cell membranes. The GLUT1 transporter has a molecular weight of approximately 55 kDa. This protein is highly expressed in erythrocytes endothelial cells lining blood vessels and in the blood-brain barrier. Its primary role is to facilitate the basal glucose uptake necessary for cellular metabolism particularly in tissues where glucose is a critical energy source.

Biological function summary

This glucose transporter plays a significant role in maintaining glucose homeostasis in the human body. GLUT1 functions independently and not as part of a complex. It ensures that glucose is available to cells with high metabolic demands including the brain and red blood cells where it remains important for survival and function. Its expression level can be influenced by various factors including hypoxia and insulin.

Pathways

GLUT1 is involved in the glycolysis and hypoxia-related pathways. It supports the glycolytic pathway by ensuring a sufficient supply of glucose to the cells which is then metabolized to produce ATP. Additionally during hypoxic conditions GLUT1 expression can increase aligning with proteins like HIF-1α which helps cells adapt by modifying their metabolism. This coordinated regulation permits cells to adjust their energy systems according to the oxygen availability.

Associated diseases and disorders

GLUT1 is implicated in glucose transporter type 1 deficiency syndrome (GLUT1 DS) and various forms of cancer. GLUT1 DS results from inadequate glucose transport into the brain presenting neurological symptoms due to energy deficiency. In cancer overexpression of GLUT1 links to increased glucose uptake and tumor growth a condition known to involve proteins like hexokinase. These associations underline GLUT1's contribution to both genetic defects and metabolic shifts in cancerous tissues.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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26 product images

Zhu, Y. et al PLoS Pathog. 2016 May 17;12(5):e1005648. doi: 10.1371/journal.ppat.1005648. eCollection 2016 May Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
GLUT1 and GLUT3 are downregulated in KSHV-infected cells in human KS tumors
Representative illustration of dual immunofluorescence detection of LANA and GLUT1 or in a normal human skin section and a Karposi Sarcoma (KS) tumor section. Tissues were fixed with paraformaldehyhde and paraffin-embedded.
Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Lanes 1 - 2: Merged signal (red and green). Green - Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free ab196357 observed at 54 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free ab196357 was shown to recognize in wild-type A549 cells as signal was lost at the expected MW in SLC2A1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SLC2A1 knockout samples were subjected to SDS-PAGE. Anti-Glucose Transporter GLUT1 antibody [EPR3915] - Low endotoxin, Azide free ab196357 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) at 1 µg/mL
Lane 1: Wild-type A549 whole cell lysate at 20 µg
Lane 2: Western blot - Human SLC2A1 (Glucose Transporter GLUT1) knockout A549 cell line (Human SLC2A1 (Glucose Transporter GLUT1) knockout A549 cell line ab261869) at 20 µg
Lane 2: Western blot - Human SLC2A1 (Glucose Transporter GLUT1) knockout A549 cell line (Human SLC2A1 (Glucose Transporter GLUT1) knockout A549 cell line ab261869)
Predicted band size: 54 kDa
Khaom, R. et al PLoS One. 2016 Aug 11;11(8):e0161163. doi: 10.1371/journal.pone.0161163. eCollection 2016 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Immunohistochemical expression of Glut1 in normal tongue epithelium and tongue cancer. Expression was greatest in lymphocytes (arrows in left upper and lower panels). In the normal oral epithelium, Glut1 was weakly expressed in the basal and spinous cells (left upper panel). In OSCC, Glut1 was upregulated, showing a level of expression comparable with lymphocytes (left and right lower panels). Scale bar, 100 μm.
Note: Glut1 = SLC2A (alternative names for the same target).
Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Glucose Transporter GLUT1 Western blot staining using rabbit Anti-Glucose Transporter GLUT1 antibody
Exposure time
Lane 1 to 2: 10 seconds
Lane 3 to 4: 30 seconds
We recommend not to boil the samples after lysis to get desired WB bands.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) at 1/50000 dilution
Lane 1: HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates unboiled at 20 µg
Lane 2: HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates boiled at 20 µg
Lane 3: 3T3-L1 (Mouse embryonic fibroblast) whole cell lysates unboiled at 20 µg
Lane 4: 3T3-L1 (Mouse embryonic fibroblast) whole cell lysates boiled at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 54 kDa
Observed band size: 40-60 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Immunofluorescence staining of HepG2 cells with purified ab115730 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab115730 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400.
Alexa Fluor^® 488 (Alexa Fluor® 488 Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab195359) and Alexa Fluor^® 647 (Alexa Fluor® 647 Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab195020) conjugated versions are available for this clone.
Flow Cytometry (Intracellular) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified ab115730 at a dilution of 1/40 (red line). The secondary antibody used was Alexa Fluorr^® 488 goat anti-rabbit at a dilution of 1/500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
Alexa Fluorr^®488 (Alexa Fluor® 488 Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab195359) and Alexa Fluorr^®647 (Alexa Fluor® 647 Anti-Glucose Transporter GLUT1 antibody [EPR3915] ab195020) conjugated versions are available for this clone.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Immunohistochemical staining of paraffin embedded human lung carcinoma with purified ab115730 at a working dilution of 1/500. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab115730 at a working dilution of 1/500. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) at 1/1000000 dilution
Lane 1: HepG2 whole cell lysate at 10 µg
Lane 2: Human fetal liver lysate at 10 µg
Lane 3: HT-29 whole cell lysate at 10 µg
Lane 4: SW480 whole cell lysate at 10 µg
Secondary
All lanes: Anti-rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 54 kDa
Observed band size: 40-60 kDa
Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
All lanes: Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) at 1/1000 dilution
Lane 1: Jurkat lysate at 10 µg
Lane 2: Mouse brain lysate at 10 µg
Lane 3: Human fetal brain lysate at 10 µg
Lane 4: 3T3L1 lysate at 10 µg
Lane 5: Human fetal liver lysate at 10 µg
Lane 6: HepG2 lysate at 10 µg
Predicted band size: 54 kDa
Flow Cytometry (Intracellular) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Overlay histogram showing HeLa cells stained with unpurified ab115730 (red line). The cells were fixed with 80% methanol (5 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 (ab115730, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Unpurified ab115730 at 1/250 dilution staining Glucose Transporter GLUT1 in Paraffin-embedded human colonic adenocarcinoma tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Unpurified ab115730 showing positive staining in human normal liver tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Unpurified ab115730 showing positive staining in human normal breast tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Unpurified ab115730 at 1/250 dilution staining Glucose Transporter GLUT1 in Paraffin-embedded human cervical carcinoma tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Unpurified ab115730 showing positive staining in human normal colon tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Unpurified ab115730 showing positive staining in human kidney carcinoma tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Unpurified ab115730 showing negative staining in human skeletal muscle tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Unpurified ab115730 showing positive staining in human urinary bladder transitional carcinoma tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Unpurified ab115730 showing negative staining in human normal heart tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
OI-RD Scanning - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Western blot: Anti-Glucose Transporter GLUT1 antibody [EPR3915] staining at 1/100000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab115730 was shown to bind specifically to Glucose Transporter GLUT1. A band was observed at 50-300 kDa in wild-type HepG2 cell lysates with no signal observed at this size in SLC2A1 knockout cell line Human SLC2A1 (Glucose Transporter GLUT1) knockout Hep G2 cell line ab280797 (knockout cell lysate ab284224). To generate this image, wild-type and SLC2A1 knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730) at 1/100000 dilution
Lane 1: Wild-type HepG2 cell lysate at 20 µg
Lane 2: LC2A1 knockout HepG2 cell lysate at 20 µg
Lane 3: A549 cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Observed band size: 50-300 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
ab115730 staining SLC2A1 in wild-type A549 cells, with negative expression in SLC2A1 knockout A549 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab115730 at 1 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
Flow Cytometry (Intracellular) - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Flow cytometry overlay histogram showing wild-type A549 (green line) and A549-SLC2A1 knockout cells stained with ab115730 (magenta line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab115730) (1x 10⁶in 100µl at 0.04 µg/ml (1/12500)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type A549 - black line, A549-SLC2A1 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in A549 WT Fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
Immunohistochemistry - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Glucose Transporter GLUT1 Immunohistochemistry staining of Human placenta using rabbit Anti-Glucose Transporter GLUT1 antibody
Immunohistochemical analysis of formalin fixed paraffin embedded human placenta labelling Glucose Transporter GLUT1 with ab115730 at a concentration of 0.05µ/ml.
The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 20mins with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5. ab115730 anti-Glucose Transporter GLUT1 was incubated at 37°C for 16mins.
Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)
Immunohistochemistry - Anti-Glucose Transporter GLUT1 antibody [EPR3915] (ab115730)
Glucose Transporter GLUT1 Immunohistochemistry staining of Human placenta using rabbit Anti-Glucose Transporter GLUT1 antibody
Immunohistochemical analysis of formalin fixed paraffin embedded human placenta labelling Glucose Transporter GLUT1 with ab115730 at a concentration of 0.2µ/ml. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20mins.
ab115730 anti-Glucose Transporter GLUT1 antibody [EPR3915] was incubated for 15mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)