Glut1 Antibody (SA0377)

Novus Biologicals | Catalog # NBP3-32385

Recombinant Monoclonal Antibody
Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # SA0377 expressed in HEK293
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Product Specifications

Immunogen

Synthetic peptide within Human GLUT1 aa 443-492 / 492. (Uniprot: P11166)

Localization

Cell membrane, Melanosome

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

54 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Glut1 Antibody (SA0377)

Glut1 Antibody (SA0377)

Immunohistochemistry: Glut1 Antibody (SA0377) [NBP3-32385]

Immunohistochemistry: Glut1 Antibody (SA0377) [NBP3-32385] - Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Glut1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32385, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Glut1 Antibody (SA0377)

Immunohistochemistry: Glut1 Antibody (SA0377) [NBP3-32385]

Immunohistochemistry: Glut1 Antibody (SA0377) [NBP3-32385] - Immunofluorescence analysis of paraffin-embedded human liver tissue labeling Glut1 with Rabbit anti-Glut1 antibody (NBP3-32385) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (NBP3-32385, green) at 1/100 dilution overnight at 4 ℃, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
Glut1 Antibody (SA0377)

Immunocytochemistry/ Immunofluorescence: Glut1 Antibody (SA0377) [NBP3-32385]

Immunocytochemistry/ Immunofluorescence: Glut1 Antibody (SA0377) [NBP3-32385] - Immunocytochemistry analysis of HeLa cells labeling Glut1 with Rabbit anti-Glut1 antibody (NBP3-32385) at 1/500 dilution.

Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Glucose Transporter GLUT1 antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
Glut1 Antibody (SA0377)

Flow Cytometry: Glut1 Antibody (SA0377) [NBP3-32385]

Flow Cytometry: Glut1 Antibody (SA0377) [NBP3-32385] - Flow cytometric analysis of Jurkat cells labeling Glut1.

Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32385, red) at 1/1,000 dilution, compared with Rabbit IgG Isotype Control (blue). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with an iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabeled sample was used as a control (cells without incubation with primary antibody; black).
Glut1 Antibody (SA0377)

Western Blot: Glut1 Antibody (SA0377) [NBP3-32385]

Western Blot: Glut1 Antibody (SA0377) [NBP3-32385] - Western blot analysis of Glut1 on different lysates with Rabbit anti-Glut1 antibody (NBP3-32385) at 1/50,000 dilution.
Lane 1: HeLa cell lysate (no heat)
Lane 2: HT-29 cell lysate (no heat)
Lane 3: HepG2 cell lysate (no heat)
Lane 4: NIH/3T3 cell lysate (no heat)
Lane 5: L-929 cell lysate (no heat)
Lane 6: Mouse brain tissue lysate (no heat)
Lane 7: Rat brain tissue lysate (no heat)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 45-60 kDa

Exposure time:
Lane 1-7 (left): 20 seconds;
Lane 1-7 (right): 1 minute 30 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/50,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.

Applications for Glut1 Antibody (SA0377)

Application
Recommended Usage

Flow Cytometry

1:500-1:1000

Immunocytochemistry/ Immunofluorescence

1:500-1:200

Immunohistochemistry-Paraffin

1:50-1:200

Western Blot

1:5000-1:50000

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Advanced Features

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Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

1*TBS (pH7.4), 0.05% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Glut1

Glucose transporter 1 (GLUT1) or solute carrier family 2 (SLC2A1) is a member of the GLUT family of monosaccharides and polyols transporters. GLUT proteins transport glucose across cellular membranes through facilitative mechanisms and play a key role in glucose homeostasis (1). Fourteen GLUT proteins have been identified in the human, which are encoded by SLC2A genes 1-14 and are broadly expressed in many cell types and tissues. GLUT family members differ in sequence homology, substrate specificity and expression patterns. Based on sequence homology, GLUT family members are classified into Class I (GLUT1, 2, 3, 4, and GLUT14), Class II (GLUT5, 7, 9, and 11), and Class III (GLUT6, 8, 10, 12 and 13) (1). Structurally, GLUT transporters are integral membrane glycoproteins consisting of 12 membrane spanning helical domains, a single N-linked glycosylation site, and having cytoplasmic facing carboxy and amino terminal domains (2).

GLUT1 (Human glycosylated form theoretical molecular weight 55kDa) functions primarily as a glucose transporter but can transport other substrates including mannose, galactose and glucosamine across the membrane (3). Like other GLUT family members, GLUT1 is broadly expressed, nevertheless it is the predominant glucose transporter expressed in red blood cells and brain endothelial cells (1). SLC2A1 mutations underscore the autosomal dominant disorder GLUT1 deficiency syndrome (GLUTI-DS) which is characterized by low glucose levels in the brain or hypoglycorrhachia due to insufficient glucose transport across the blood brain barrier (2, 4, 5). Phenotypically, GLUT1-DS is characterized by early onset seizures, neurologic developmental delay, microcephaly, and ataxia (4). GLUT1 is highly expressed in the endothelium of cutaneous vascular lesions and serves as a marker for the diagnosis of juvenile or infantile hemangiomas (6).

References

1. Augustin, R. (2010). The protein family of glucose transport facilitators: It's not only about glucose after all. IUBMB Life. https://doi.org/10.1002/iub.315

2. Mueckler, M., & Thorens, B. (2013). The SLC2 (GLUT) family of membrane transporters. Molecular Aspects of Medicine. https://doi.org/10.1016/j.mam.2012.07.001

3. Stein, W. D., & Litman, T. (2015). Carrier-Mediated Transport. In Channels, Carriers, and Pumps. https://doi.org/10.1016/b978-0-12-416579-3.00004-6

4. Pearson, T. S., Akman, C., Hinton, V. J., Engelstad, K., & De Vivo, D. C. (2013). Phenotypic spectrum of glucose transporter type 1 deficiency syndrome (Glut1 DS). Current Neurology and Neuroscience Reports. https://doi.org/10.1007/s11910-013-0342-7

5. Messana, T., Russo, A., Vergaro, R., Boni, A., Santucci, M., & Pini, A. (2018). Glucose transporter type 1 deficiency syndrome: Developmental delay and early-onset ataxia in a novel mutation of the SLC2A1 gene. Journal of Pediatric Neurosciences. https://doi.org/10.4103/JPN.JPN_169_17

6. van Vugt, L. J., van der Vleuten, C. J. M., Flucke, U., & Blokx, W. A. M. (2017). The utility of GLUT1 as a diagnostic marker in cutaneous vascular anomalies: A review of literature and recommendations for daily practice. Pathology Research and Practice. https://doi.org/10.1016/j.prp.2017.04.023

Long Name

Glucose Transporter Type 1

Alternate Names

DYT17, DYT18, DYT9, EIG12, GLUT1DS, SLC2A1

Gene Symbol

SLC2A1

Additional Glut1 Products

Product Documents for Glut1 Antibody (SA0377)

Certificate of Analysis

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Product Specific Notices for Glut1 Antibody (SA0377)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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