Glut1 Overexpression Lysate

Name: Glut1 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-10406

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-10406

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Product Datasheet / COA / SDS

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Glut1 Overexpression Lysate

Glut1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC222696

Accession#: NM_006516

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

54.1 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.

Type

Overexpression

Scientific Data Images for Glut1 Overexpression Lysate

Western Blot: Glut1 Overexpression Lysate [NBP2-10406]

Western Blot: Glut1 Overexpression Lysate [NBP2-10406] - GLUT1 Overexpression Lysate (Adult Normal) [NBP2-10406] Left lane: Empty vector transfected control (HEK293 cell lysate); Right lane: Over-expression lysate for GLUT1.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. 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 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for Glut1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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