Anti glp 1r

Manufactured by Abcam

Sourced in United States, United Kingdom

Anti-GLP-1R is a laboratory reagent used for the detection and quantification of the glucagon-like peptide-1 receptor (GLP-1R) in various biological samples. It is a specific antibody that binds to the GLP-1R protein, allowing researchers to study the expression and localization of this receptor in cells and tissues.

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7 protocols using anti glp 1r

Western Blot Analysis of Protein Expressions

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Protein expression was examined by Western blot analysis. In brief, the cells were lysed in RIPA buffer and the total protein was separated by SDS-PAGE. Primary antibodies used in this study include: anti-adiponectin antibody (CST, Cat. No. 2789, dilution 1:500), anti-Foxo-1 antibody (CST, Cat. No. 2880, dilution 1:500), anti-p-Foxo-1 antibody (CST, Cat. No. 2486, dilution 1:500), anti-GLP-1R (Abcam, Cat No. ab39072, dilution 1:1000), anti-HA tag (Abcam, Cat No. ab18181, dilution 1:1000), and anti-GAPDH (CST, Cat. No. 5174, dilution 1:5000). HRP-conjugated goat-anti-rabbit secondary antibody was obtained from Tiandeyue (Cat. No.S001), and diluted by 1:20000 in the Western blot analysis. For each Western blot analysis, 3 independent experiments were carried out and representative results are shown in the figures.

Wang A., Li T., An P., Yan W., Zheng H., Wang B, & Mu Y. (2017). Exendin-4 Upregulates Adiponectin Level in Adipocytes via Sirt1/Foxo-1 Signaling Pathway. PLoS ONE, 12(1), e0169469.

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GLP-1R Protein Expression Analysis

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The total protein of 30 μg was fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and then transferred to polyvinylidene fluoride membranes. Primary antibodies used in Western blotting were as follows: anti-GLP-1R (1:1000, ab218532, Abcam, UK), Actin antibody(I-19): sc-1616 HRP (B1815, Santa Cruz, USA). Secondary antibodies were horseradish peroxidase-conjugated goat anti-rabbit lgG (1:20000 Santa Cruz, USA). The results were visualized using an enhanced chemiluminescence system (ECL^™ prime, Amersham, UK). Densitometric analysis was conducted using ImageJ software (NIH, USA).

Kajitani Y., Miyazawa T., Inoue T., Kajitani N., Fujita M., Takeichi Y., Miyachi Y., Sakamoto R, & Ogawa Y. (2023). High frequency of germline recombination in Nestin-Cre transgenic mice crossed with Glucagon-like peptide 1 receptor floxed mice. PLOS ONE, 18(12), e0296006.

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Protein Expression Analysis of Pancreatic Cells

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Cultured islets or Min6 cells were washed in PBS and lysed. PVDF membranes were incubated with anti-TCF7L2 (#2565), anti-actin (#4967), anti-p-AKT (Serine473, #9271), anti-AKT (#9272), anti-p-GSK3β (Ser9 #9336), anti-PARP (#9542), anti-GAPDH (#2118), anti-c-casp3 (#9661), anti-stat3 (#9132), anti-p-stat3 (Tyr705, #9131), anti-PKA C-α ( #5842; all from Cell Signaling, Danvers, MA, USA), anti-β-catenin (ab6302), anti-GLP-1R (ab39072), anti-p-Jak2 (ab68268; all from Abcam), followed by incubation with horseradish-peroxidase-linked IgG peroxidase. The bands were visualized and densities of the bands were analyzed using Tanon ChemImaging Systems (Nanjing, China).

Yao D.D., Yang L., Wang Y., Liu C., Wei Y.J., Jia X.B., Yin W, & Shu L. (2015). Geniposide promotes beta-cell regeneration and survival through regulating β-catenin/TCF7L2 pathway. Cell Death & Disease, 6(5), e1746-.

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Western Blot Analysis of Nuclear and Total Cell Proteins

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For immunodetection of p65, p–cAMP-response element binding protein (CREB), CREB, and GLP-1R, 30 μg of protein from nuclear or total cell extracts were separated using SDS-PAGE and transferred onto a polyvinylidene difluoride membrane (Boehringer Mannheim, Mannheim, Germany). Western blot was performed as previously described.^{9 (link)} Membranes were probed with rabbit polyclonal anti-p65, anti-CREB, anti-phosphoCREB (1:1000, all from Cell Signaling Technology, Beverly, MA, USA) and anti-GLP-1R (1:300, Abcam, Cambridge, UK). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, 1:10,000; Sicgen, Cantanhede, Portugal) and Lamin B (1:4000, Abcam) were used as loading controls for total and nuclear extracts, respectively. Immunoreactive bands were detected by enhanced chemiluminescence (ECL) substrate using an imaging system (VersaDoc 4000 MP; Bio-Rad) and quantification was performed using ImageJ software (version 1.47, http://imagej.nih.gov/ij/; provided in the public domain by the National Institutes of Health, Bethesda, MD, USA).

Gonçalves A., Lin C.M., Muthusamy A., Fontes-Ribeiro C., Ambrósio A.F., Abcouwer S.F., Fernandes R, & Antonetti D.A. (2016). Protective Effect of a GLP-1 Analog on Ischemia-Reperfusion Induced Blood–Retinal Barrier Breakdown and Inflammation. Investigative Ophthalmology & Visual Science, 57(6), 2584-2592.

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Neuroprotective Effects of Bifidobacterium lactis NJ241

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Bifidobacterium animalis subsp. lactis NJ241 (CGMCC NO.20816) was obtained from Thankcome Biotechnology Co., Ltd (Suzhou, China). MPTP was purchased from Med-ChemExpress (Shanghai, China). The antibodies utilized in this study were obtained as follows: rabbit anti-TH, anti-GLP-1R, anti-GFAP, and mouse anti-GAPDH polyclonal antibodies were acquired from Proteintec (Wuhan, China); rabbit anti-Iba-1, anti-PGC-1α, and anti-GLP-1R monoclonal antibodies were sourced from Abcam (Shanghai, China); rabbit anti-ZO-1 polyclonal antibody was obtained from Servicebio (Wuhan, China); rabbit anti-occludin polyclonal antibody was acquired from Wanleibio (Shenyang, China); and rabbit anti-GLP-1 polyclonal antibody, Alexa Fluor 594-conjugated goat anti-rabbit IgG, HRP-conjugated goat anti-mouse IgG, and HRP-conjugated goat anti-rabbit IgG were obtained from Abways Technology (Shanghai, China).

Dong Y., Qi Y., Chen J., Han S., Su W., Ma X., Yu Y, & Wang Y. (2024). Neuroprotective Effects of Bifidobacterium animalis subsp. lactis NJ241 in a Mouse Model of Parkinson's Disease: Implications for Gut Microbiota and PGC-1α. Molecular neurobiology.

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GLP-1R Expression After Intracerebral Hemorrhage

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Mice were euthanized at 3 days after ICH and brain homogenate was extracted from the entire right brain hemisphere of each mouse. Western blot was performed as described previously^{12 (link)}. Briefly, total protein was extracted using RIPA buffer with 1% PMSF (Solarbio, Beijing, China). After quantitative analysis of the total protein concentration with BCA method, the samples were separated by 10% SDS-PAGE and transferred to PVDF membrane (Millipore, USA). To separate the region where the target protein will appear, the membrane was cut along the molecular weight marker. Then the PVDF membrane was blocked with Blocking Buffer (Beyotime, Nanjing, China) at room temperature for 1 h and incubated at 4 °C on a shaker with the anti-GLP-1R (1:1000, Abcam, Cambridge, MA, USA)^{37 (link)} and anti-GAPDH (1:2000, Servicebio, China) antibodies. After washing with TBST, the membrane was placed in the configured secondary antibodies (Goat anti-mouse/Rabbit HRP, 1:5000), incubated at room temperature for 1 h. The immunoblot was visualized with Ultra High Sensitivity ECL Kit (GlpBio, Montclair, USA) through Amersham Imager 600. The relative intensity of the bands was measured by ImageJ (NIH, USA).

Zhang Y., Liu Y., Yong V.W, & Xue M. (2023). Omarigliptin inhibits brain cell ferroptosis after intracerebral hemorrhage. Scientific Reports, 13, 14339.

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Westernblot Analysis of Cardiac Proteins

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Both H9C2 cells and myocardial tissues were harvested for Western blot analysis (Western Blot Detection Kit, Elabscience) following standard protocol according to the manufacturer’s instructions and Towbin system buffer was used. Primary antibodies used in Western blot were as follows: anti-GLP-1R (1:200, Abcam, UK), anti-Akt (1:200, Abcam, UK), anti-p-Akt (1:200, Abcam, UK), anti-PI3K (1:500, Santa Cruz, USA), anti-PKC (1:500, Abcam, UK), anti-PKCα (1:500, Abcam, UK), anti-PKCβ (1:500, Abcam, UK), anti-PKCγ (1:500, Abcam, UK), anti-PKCδ (1:500, Abcam, UK) and anti-β-actin (1:500, Santa Cruz, USA). Secondary antibodies were horseradish peroxidase-conjugated goat anti-rabbit lgG (Santa Cruz, USA) at 1:5000 dilution. The results were visualized using an enhanced chemiluminescence system (ECL, Amersham). Densitometric analysis was conducted using ImageJ software (NIH, USA).

Pan X., Chen J., Wang T., Zhang M., Wang H, & Gao H. (2019). Essential Role Of High Glucose-Induced Overexpression Of PKCβ And PKCδ In GLP-1 Resistance In Rodent Cardiomyocytes. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy, 12, 2289-2302.

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