Anti cb1

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Anti-CB1 is a laboratory equipment designed for research purposes. It functions as a specific antibody that targets the CB1 receptor, which is involved in various physiological processes. This product is intended for use in research applications that require the detection or study of the CB1 receptor.

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3 protocols using anti cb1

Characterization of CXCR4 and Cannabinoid Signaling

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Brilliant violet 421 anti-human CXCR4, allophycocyanin anti-human CD5 and phycoerythrin anti-human CD19 antibodies were obtained from BioLegend, BD Biosciences and Dako, respectively. Antibodies used for Western blotting: anti-phospho ERK1/2 (Thr202/Tyr204), anti-total ERK1/2, anti-phospho p38 (Thr180/Tyr182), anti-total p38, anti-phospho Akt (Ser473) and anti-total Akt were all from Cell Signaling Technology (Danvers, MA, USA). Antibody anti-CB1 was purchased from ThermoFisher (cat#PA1-743), and anti-CB2 was from Santa-Cruz (cat#293188). CXCL12 was from R&D Systems (biotechne, Abingdon, UK), 2-arachidonoylglycerol was from Tocris Bioscience and Calcein-AM was purchased from ThermoFisher Scientific. The CB1 antagonist AM6545 was purchased from Tocris Bioscience and the CB1 inverse agonist SR141716 and the CB2 antagonist SR144528 were from Cayman Chemical. The CXCR4 antagonist AMD3100 was purchased from Sigma.

Merrien M., Wasik A.M., Melén C.M., Morsy M.H., Sonnevi K., Junlén H.R., Christensson B., Wahlin B.E, & Sander B. (2023). 2-Arachidonoylglycerol Modulates CXCL12-Mediated Chemotaxis in Mantle Cell Lymphoma and Chronic Lymphocytic Leukemia. Cancers, 15(5), 1585.

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Western Blot Analysis of Protein Signaling

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After centrifugation and supernatant discard, the proteins were extracted from the pellet using RIPA Buffer and quantified using Bradford Assay (Bio-Rad, Hercules, CA, USA). After denaturation at 95 °C, 25 µg of protein for each samples underwent separation using SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and then they were transferred on PVDF membrane (Immobilon–P, Millipore, Burlington, MA, USA). Then, 5% skimmed milk in TBS was used for blocking at room temperature for 1 hour, followed by the overnight incubation at 4 °C. The antibodies used were: Anti-CB1 (1:500, ThermoFisher Scientific, Rockford, IL, USA), anti-GAPDH HRP conjugate (1:1000, Cell Signaling, Danvers, MA, USA), anti-p-Akt (1:1000; Cell Signaling, Danvers, MA, USA), anti-Akt (1:1000, Cell Signaling, Danvers, MA, USA), anti-p-ERK1/2 (1:2000; Cell Signaling, Danvers, MA, USA) and anti-ERK2 (1:1000; Cell Signaling, Danvers, MA, USA). The antibody mouse anti-rabbit IgG-HRP (1:2000, Santa Cruz Biotechnology, Dallas, TX, USA) was used as a secondary antibody to be incubated with the membranes for 1 h at room temperature. In order to acquire the bands, ChemiDoc™ MP System (Bio-Rad) was after exposition to enhanced chemiluminescence system (Luminata Western HRP Substrates, Millipore, Burlington, MA, USA).

Blando S., Raffaele I., Chiricosta L., Valeri A., Gugliandolo A., Silvestro S., Pollastro F, & Mazzon E. (2022). Cannabidiol Promotes Neuronal Differentiation Using Akt and Erk Pathways Triggered by Cb1 Signaling. Molecules, 27(17), 5644.

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Western Blot Analysis of HSC Markers

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After various treatment, HSCs were washed twice with PBS and scraped in a lysis buffer (20 mM Tris-HCl, 250 mM sucrose, pH 7.2) containing 1X protease inhibitor mixture (Roche, Indianapolis, IN). Proteins were denatured with SDS buffer and boiled for 5 min. Samples were run by SDS-PAGE, transferred onto a PVDF membrane, and blocked using 5% nonfat dry milk (NFDM) in Tris-buffered saline with 0.1% Tween 20 (TBST). Then, the membranes were probed with the following primary antibodies overnight at 4 °C: anti-α-SMA (1:1000, Abcam, Cambridge, MA), anti-desmin (1:500, BD Pharmingen, San Jose, CA), anti-CB1 (1:1000, Thermo, Waltham, MA), anti-CB2 (1:1000, Abcam), anti-TIMP-1 (1:500, Santa Cruz, Dallas, TX), anti-collagen I (1:1000, Calbiochem, Billerica, MA), anti-Nox1 (1:500, Abcam), anti-Nox4 (1:500, Santa Cruz), anti-Nrf2 (1:1000, Santa Cruz), and anti-β-actin (1:10000, Santa Cruz). After washing with TBST three times, membranes were incubated with the following secondary antibodies for 1 h at room temperature, respectively with corresponding goat anti-rabbit-HRP or rabbit anti-mouse-HRP (1:10000, Santa Cruz) based on animals used for raising primary antibody. After washing for the final three times, bands were detected by the addition of a chemiluminescent substrate and visualized on Kodak Omat X-ray film.

Wang M., Abais J.M., Meng N., Zhang Y., Ritter J.K., Li P.L, & Tang W.X. (2014). Upregulation of cannabinoid receptor-1 and fibrotic activation of mouse hepatic stellate cells during Schistosoma J. infection: Role of NADPH oxidase. Free radical biology & medicine, 71, 109-120.

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