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71831
Glut1 (IHC404) Mouse mAb
Primary Antibodies
Monoclonal Antibody

Glut1 (IHC404) Mouse mAb #71831

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  1. IHC
Immunohistochemistry Image 1: Glut1 (IHC404) Mouse mAb
Immunohistochemical analysis of paraffin-embedded Hep G2 cell pellet (left, high-expressing) or MOLT-4 cell pellet (right, low-expressing) using Glut1 (IHC404) Mouse mAb.
Immunohistochemistry Image 2: Glut1 (IHC404) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Glut1 (IHC404) Mouse mAb.
Immunohistochemistry Image 3: Glut1 (IHC404) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human oral squamous cell carcinoma using Glut1 (IHC404) Mouse mAb.
Immunohistochemistry Image 4: Glut1 (IHC404) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of the tonsil using Glut1 (IHC404) Mouse mAb.
Immunohistochemistry Image 5: Glut1 (IHC404) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Glut1 (IHC404) Mouse mAb.
Immunohistochemistry Image 6: Glut1 (IHC404) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human neuroendocrine lung carcinoma using Glut1 (IHC404) Mouse mAb.
Immunohistochemistry Image 7: Glut1 (IHC404) Mouse mAb
Immunohistochemical analysis of paraffin-embedded normal human lung using Glut1 (IHC404) Mouse mAb.
Immunohistochemistry Image 8: Glut1 (IHC404) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin lymphoma using Glut1 (IHC404) Mouse mAb.
Immunohistochemistry Image 9: Glut1 (IHC404) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Glut1 (IHC404) Mouse mAb (left) compared to concentration-matched Mouse (E5Y6Q) mAb IgG2a Isotype Control #61656 (right).
Immunohistochemistry Image 10: Glut1 (IHC404) Mouse mAb
Immunohistochemical analysis of paraffin-embedded normal human testis using Glut1 (IHC404) Mouse mAb.
To Purchase # 71831S
Product # Size Price
71831S
100 µl $ 285

Supporting Data

REACTIVITY H
SENSITIVITY Endogenous
MW (kDa)
Source/Isotype Mouse IgG2a

Application Key:

  • WB-Western Blot
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Vir-Virus
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Usage Information

Application Dilution
Immunohistochemistry (Paraffin) 1:1600 - 1:3200

Storage

Supplied in a Tris-based buffer with 1% BSA and less than 0.1% sodium azide. Stable for 24 months when stored at 4°C. Do not aliquot the antibody.

Protocol

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Immunohistochemistry (Paraffin)

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. Xylene.
  2. Ethanol, anhydrous denatured, histological grade (100% and 95%).
  3. Deionized water (dH2O).
  4. Hematoxylin (optional).
  5. Wash Buffer:
    1. 1X Tris Buffered Saline with Tween® 20 (TBST): To prepare 1L 1X TBST add 100 ml 10X Tris Buffered Saline with Tween® 20 (#9997) to 900 ml dH20, mix.
  6. SignalStain® Antibody Diluent (#8112).
  7. 1X Citrate Unmasking Solution: To prepare 250 mL of 1X citrate unmasking solution, dilute 25 ml of SignalStain® Citrate Unmasking Solution (10X) (#14746) with 225 mL of dH2O.
  8. 3% Hydrogen Peroxide: To prepare 100 ml, add 10 ml 30% H2O2 to 90 ml dH2O.
  9. Blocking Solution: TBST/5% Normal Goat Serum or 1X Animal-Free Blocking Solution.
    1. TBST/5% Normal Goat Serum: to 5 ml 1X TBST, add 250 µl Normal Goat Serum (#5425).
    2. 1X Animal-Free Blocking Solution: to 4 mL of dH2O add 1 ml of Animal-Free Blocking Solution (5X) (#15019).
  10. Detection System: SignalStain® Boost IHC Detection Reagents (HRP, Mouse #8125).
  11. Substrate: SignalStain® DAB Substrate Kit (#8059).
  12. Hematoxylin: Hematoxylin (#14166).
  13. Mounting Medium: SignalStain® Mounting Medium (#14177).

B. Deparaffinization/Rehydration

NOTE: Do not allow slides to dry at any time during this procedure.

  1. Deparaffinize/hydrate sections:
    1. Incubate sections in three washes of xylene for 5 min each.
    2. Incubate sections in two washes of 100% ethanol for 10 min each.
    3. Incubate sections in two washes of 95% ethanol for 10 min each.
  2. Wash sections two times in dH2O for 5 min each.

C. Antigen Unmasking

For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.

D. Staining

  1. Wash sections in dH2O three times for 5 min each.
  2. Incubate sections in 3% hydrogen peroxide for 10 min.
  3. Wash sections in dH2O two times for 5 min each.
  4. Wash sections in wash buffer for 5 min.
  5. Block each section with 100–400 µl of preferred blocking solution for 1 hr at room temperature.
  6. Remove blocking solution and add 100–400 µl primary antibody diluted in SignalStain® Antibody Diluent (#8112) to each section. Incubate overnight at 4°C.
  7. Equilibrate SignalStain® Boost Detection Reagent (HRP, Mouse #8125) to room temperature.
  8. Remove antibody solution and wash sections with wash buffer three times for 5 min each.
  9. Cover section with 1–3 drops SignalStain® Boost Detection Reagent (HRP, Mouse #8125) as needed. Incubate in a humidified chamber for 30 min at room temperature.
  10. Wash sections three times with wash buffer for 5 min each.
  11. Add 1 drop (30 µl) SignalStain® DAB Chromogen Concentrate to 1 ml SignalStain® DAB Diluent and mix well before use.
  12. Apply 100–400 µl SignalStain® DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity.
  13. Immerse slides in dH2O.
  14. If desired, counterstain sections with hematoxylin (#14166).
  15. Wash sections in dH2O two times for 5 min each.
  16. Dehydrate sections:
    1. Incubate sections in 95% ethanol two times for 10 sec each.
    2. Repeat in 100% ethanol, incubating sections two times for 10 sec each.
    3. Repeat in xylene, incubating sections two times for 10 sec each.
  17. Mount sections with coverslips and mounting medium (#14177).

DETECTION REAGENT/SUBSTRATE COMPATIBILITY
RECOMMENDED
DETECTION REAGENTS
SignalStain® Boost IHC Detection Reagent (HRP, Mouse) #8125 SignalStain® Boost IHC Detection Reagent (AP, Mouse) #31926
COMPATIBLE
CHROMOGEN
SignalStain® DAB Substrate Kit #8059 SignalStain® Vibrant Red Alkaline Phosphatase Substrate Kit #76713
SignalStain® Vivid Purple Peroxidase Substrate Kit #96632  

NOTE: Use of detection reagents other than those specified in this protocol may require further optimization of the primary antibody to account for the different sensitivities of the detection reagents.


posted February 2010

revised June 2020

Protocol Id: 280

Specificity / Sensitivity

Glut1 (IHC404) Mouse mAb recognizes endogenous levels of total Glut1 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the C terminal of Glut1 protein.

Background

Glucose transporter 1 (Glut1, SLC2A1) is a widely expressed transport protein that displays a broad range of substrate specificity in transporting a number of different aldose sugars as well as an oxidized form of vitamin C into cells (1,2). Glut1 is responsible for the basal-level uptake of glucose from the blood through facilitated diffusion (2). Research studies show that Glut1 and the transcription factor HIF-1α mediate the regulation of glycolysis by O-GlcNAcylation in cancer cells (3). Additional studies demonstrate that Glut1 is required for CD4 T cell activation and is critical for the expansion and survival of T effector (Teff) cells (4). Mutations in the corresponding SLC2A1 gene cause GLUT1 deficiency syndromes (GLUT1DS1, GLUT1DS2), a pair of neurologic disorders characterized by delayed development, seizures, spasticity, paroxysmal exercise-induced dyskinesia, and acquired microcephaly (5,6). Two other neurologic disorders - dystonia-9 (DYT9) and susceptibility to idiopathic generalized epilepsy 12 (EIG12) - are also caused by mutations in the SLC2A1 gene (7,8).
  1. Ferrer, C.M. et al. (2014) Mol Cell 54, 820-31.
  2. Deng, D. et al. (2014) Nature 510, 121-5.
  3. Agus, D.B. et al. (1997) J Clin Invest 100, 2842-8.
  4. Macintyre, A.N. et al. (2014) Cell Metab 20, 61-72.
  5. Wang, D. et al. (2005) Ann Neurol 57, 111-8.
  6. Schneider, S.A. et al. (2009) Mov Disord 24, 1684-8.
  7. Weber, Y.G. et al. (2011) Neurology 77, 959-64.
  8. Suls, A. et al. (2009) Ann Neurol 66, 415-9.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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