Anti glut4 antibody

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The Anti-GLUT4 antibody is a research tool used to detect and quantify the expression of the GLUT4 glucose transporter protein. GLUT4 is a key regulator of glucose homeostasis and is primarily expressed in adipose tissue and skeletal muscle. This antibody can be utilized in techniques such as Western blotting, immunohistochemistry, and flow cytometry to study GLUT4 distribution and function.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Phospho irs 1ser307, by Cell Signaling Technology (1 mentions) Protease and phosphatase inhibitor cocktails, by Wuhan Servicebio Technology (1 mentions) Hrp conjugated goat anti mouse, by Cell Signaling Technology (1 mentions) Hrp conjugated goat anti rabbit antibody, by Cell Signaling Technology (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) 3 isobutyl 1 methylxanthine, by Merck Group (1 mentions) Oil red o, by Merck Group (1 mentions) Phospho akt ser473, by Cell Signaling Technology (1 mentions) Insulin, by Merck Group (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions) Mouse tnf α elisa kit, by Neobioscience (1 mentions) Rat insulin radioimmunoassay kit, by Merck Group (1 mentions) Protein g agarose bead, by Santa Cruz Biotechnology (1 mentions) Enhanced chemiluminescence ecl kit, by Cytiva (1 mentions) Doc2b antibody, by Abcam (1 mentions) Goat anti rabbit horseradish peroxidase, by Bio-Rad (1 mentions) Humulin r, by Eli Lilly (1 mentions)

5 protocols using anti glut4 antibody

4-HIL Signaling Pathway Analysis

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4-HIL (>98% purity) was obtained from Sigma-Aldrich (St Louis, MO, USA), dissolved in triple-distilled water at a concentration of 100 mM, stored at −20°C, and then further diluted in cell culture medium immediately before use.
Antibodies against TACE and TIMP3 were purchased from Abcam (Cambridge, UK). Antibodies against IRS-1, IRS-2, phospho-IRS-1 Ser³⁰⁷, phospho-IRS-1 Ser³¹⁸, Akt, and phospho-Akt Ser⁴⁷³ were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-GLUT4 antibody was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Goat anti-mouse HRP-conjugated and goat anti-rabbit HRP-conjugated antibodies were also purchased from Cell Signaling Technology. The mouse TNF-α ELISA kit was obtained from Neobioscience (Shenzhen, People’s Republic of China).
Penicillin, streptomycin, Dulbecco’s Modified Eagle’s Medium (DMEM), and fetal bovine serum were obtained from GIBCO BRL Life Technologies (Grand Island, NY, USA). Insulin, 3-isobutyl-1-methylxanthine, dexamethasone, oil red O, and 2-deoxy-[³H]-d-glucose (2-DOG) were purchased from Sigma-Aldrich. Protease and phosphatase inhibitor cocktails were purchased from Servicebio (Wuhan Goodbio Technology CO., Ltd, Wuhan, People’s Republic of China). The other reagents used in this study were obtained from Sigma-Aldrich.

Gao F., Du W., Zafar M.I., Shafqat R.A., Jian L., Cai Q, & Lu F. (2015). 4-Hydroxyisoleucine ameliorates an insulin resistant-like state in 3T3-L1 adipocytes by regulating TACE/TIMP3 expression. Drug Design, Development and Therapy, 9, 5727-5736.

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Antibody Sources for Protein Analysis

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Rabbit anti-Munc18c and rabbit anti-Syn4 antibodies were generated in-house as described [17 (link), 32 (link)]. The rabbit anti-Syn4 antibody used for immunoprecipitation was purchased from Chemicon (Temecula, CA, USA). SNAP23 and SNAP25 antibodies were purchased from Affinity Bioreagents (Golden, CO, USA). The Doc2b antibody was purchased from Abcam (Cambridge, MA, USA). The Munc18-1 and VAMP2 antibodies were acquired from Synaptic Systems (Gottingen, Germany). The Syn1A antibody was obtained from Sigma (St Louis, MO, USA). The Akt and phosphoserine (Ser 473)-specific Akt antibodies were purchased from Cell Signaling (Beverley, MA, USA). Goat anti-rabbit–horseradish peroxidase and anti-mouse– horseradish peroxidase secondary antibodies were purchased from Bio-Rad (Hercules, CA, USA). Protein G+ agarose beads and anti-GLUT4 antibody were acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The rat insulin radioimmunoassay kit was acquired from Millipore (Billerica, MA, USA). The enhanced chemiluminescence (ECL) kit was purchased from Amersham Biosciences (Pittsburgh, PA, USA). Humulin R was obtained from Eli Lilly (Indianapolis, IN, USA).

Ramalingam L., Oh E, & Thurmond D.C. (2014). Doc2b enrichment enhances glucose homeostasis in mice via potentiation of insulin secretion and peripheral insulin sensitivity. Diabetologia, 57(7), 1476-1484.

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AMPK Activation and GLUT4 Translocation

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Anti-Phos-AMPKα-Thr172 antibody and anti-AMPKα antibody were purchased from the U.S. Cell Signaling, Inc.; anti-GLUT4 antibody was obtained from Santa Cruz Biotechnology; anti-α-tubulin antibody was obtained from Merck Millipore, Billerica, MA; lipopolysaccharide (LPS, Escherichia coli 0111: B4) was purchased from USA Sigma Company; insulin kit was purchased from the U.S. Adlitteram Diagnostic Laboratories Inc.; Membrane Protein Extraction Kit was purchased from the Fermentas International Inc.

Zheng X., Xu M, & Fang Q. (2014). Role of AMPKα in Skeletal Muscle Glycometabolism Regulation and Adaptation in relation to Sepsis. BioMed Research International, 2014, 390760.

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Myostatin and Insulin Signaling Assay

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Recombinant myostatin protein was obtained from R&D System (Minneapolis, MN). Recombinant insulin protein was purchased from MBL International Co. (Woburn MA). Antibodies against phospho‐IRS‐1 (Tyr895), phospho‐PI3K, phospho‐GSK3β (Ser9), phosphor‐AKT (Ser473 & Thr308), phospho‐AMPK (Thr172), and endogenous IRS‐1, p85‐PI3K, p55‐PI3K, AKT, GSK3β, AMPK, and AICAR were purchased from Cell Signaling Technology (Beverly, MA). Anti‐Glut4 antibody was from Santa Cruz (Santa Cruz, CA). Antibodies against PGC1α and β‐tubulin were obtained from AbCam Inc. (Cambridge, MA). SB431542 was from Cayman Chem. Inc. (Ann Arbor, MI).

Liu X., Bauman W.A, & Cardozo C.P. (2018). Myostatin inhibits glucose uptake via suppression of insulin‐dependent and ‐independent signaling pathways in myoblasts. Physiological Reports, 6(17), e13837.

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Evaluating Insulin Responsiveness in MSCs

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To evaluate whether MSCs were responsive to insulin, glucose uptake and the cellular localization of GLUT4 were first evaluated in MSCs not treated with GCs (Exp 1 and 2) from T1 to T6.
For the glucose uptake assay, 3x10³ cells/well were plated in a 96-well plate and treated according to the above protocol; after insulin stimulation, 10 mM of 2-deoxyglucose (2-DG) was added for 20 minutes, and a colorimetric assay was performed following the manufacturer’s instructions. The readings were at 420 nm in a microplate reader (Thermo Scientific Multiskan GO Microplate Spectrophotometer, Milano, Italy).
For the cellular distribution of GLUT4, 1.5x10⁴ cells (Exp 1 and 2 derived from the 7 patients) were seeded in triplicate on coverslips and treated as indicated before until T5 sampling time. Cells were then washed, fixed with 4% PFA and permeabilized for 30 min. Subsequently, cells were incubated with anti-GLUT4 antibody (Santa Cruz Biotechnology, USA) followed by treatment for 30 min with a goat anti-mouse FITC-conjugated antibody (23 (link)). Finally, coverslips were mounted on glass slides in Vectashield (Vectorlabs, CA, USA), and confocal imaging was performed using a Zeiss LSM510/Axiovert 200 M microscope with an objective lens at 20× magnification (24 (link)). Line scans were acquired excluding nuclear regions, and GLUT4 immunofluorescence was analyzed as described elsewhere.

Di Vincenzo M., Martino M., Lariccia V., Giancola G., Licini C., Di Benedetto G., Arnaldi G, & Orciani M. (2022). Mesenchymal Stem Cells Exposed to Persistently High Glucocorticoid Levels Develop Insulin-Resistance and Altered Lipolysis: A Promising In Vitro Model to Study Cushing’s Syndrome. Frontiers in Endocrinology, 13, 816229.

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