Anti β2 ar

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Anti-β2-AR is a laboratory research product that specifically binds and detects the beta-2 adrenergic receptor (β2-AR) protein. It can be used in various research applications involving the identification and analysis of the β2-AR target.

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7 protocols using anti β2 ar

Macrophage Polarization Pathway Analysis

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Phorbol-12-myristate-13-acetate (PMA) was bought from Sigma Aldrich (St. Louis, Missouri, USA), human IL-4 and human IL-13 were purchased from Peprotech (200-04 and 200-13, Rocky Hill, USA).The anti-CD163, anti-CD206, anti-β2-AR and anti-PKA antibodies were purchased from Santa Cruz Biotechnology(sc-33559, sc-58986, sc-271322 and sc-98951, Ca, USA). Phenylmethylsulfonyl fluoride (PMSF) was bought from beyotime biotechnology (Shanghai, China). Primary antibodies against STAT3, pSTAT3, pCREB were purchased from Cell Signaling Technology (4904s, 9131s and 9198s, Danvers, MA, USA). Anti-CREB antibody was purchased from Abcam (ab32515, Cambridge, MA, USA). AlexaFluor488 donkey anti-mouse antibody and AlexaFluor594 goat anti-rabbit antibody were purchased from ThermoFisher Scientific (A11037, A21202, Waltham, MA, USA). Fluorescein isothiocyanate dextran (FITC-dextran) was from Sigma Aldrich (FD40s, St. Louis, Missouri, USA). AlexFluor 647 was from Fcmacs (FMS-Msaf64701, Nanjing, Jiangsu, China).

Wu J.J., Yang Y., Peng W.T., Sun J.C., Sun W.Y, & Wei W. (2019). G protein-coupled receptor kinase 2 regulating β2-adrenergic receptor signaling in M2-polarized macrophages contributes to hepatocellular carcinoma progression. OncoTargets and therapy, 12, 5499-5513.

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Quantifying Neuropathic Receptor Profiles

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DRGs (T13–L2) from AMS-treated control or NMD rats were dissected out to assess the expression of P2X1/2/3Rs and β₂-ARs. The antibodies used were anti-GAPDH (1:2000, Goodhere, Hangzhou, China), anti-P2X1/2/3R (1:1000, Alomone Labs, Hadassah Ein Kerem, Israel) and anti-β₂-AR (1:500, Santa Cruz Biotechnology, Texas, USA). Band density was measured using ImageJ software. The P2X1/2/3R and β₂-AR expression was normalized to GAPDH.

Hu S., Sun Q., Du W.J., Song J., Li X., Zhang P.A., Xu J.T, & Xu G.Y. (2020). Adult Stress Promotes Purinergic Signaling to Induce Visceral Pain in Rats with Neonatal Maternal Deprivation. Neuroscience Bulletin, 36(11), 1271-1280.

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Isoproterenol and Trichostatin A Modulate Cell Signaling

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Isoproterenol and Trichostatin A (TSA) were purchased from Sigma Aldrich (St. Louis, MO, USA) and used at 10 µM and 100 nM, respectively. Murine TNF-α was a gift from the VIB Department for Molecular Biomedical Research of Ghent University (VIB-UGent, Gent, Belgium) and was used at 2000 IU/ml. Insulin-like growth factor-1 (IGF-1) was from ImmunoTools (Friesoythe, Germany) and was used at 10 ng/ml. Anti-β₂-AR, anti-TNF-R1, anti-myogenin, anti-PARP, anti-P-H3-Ser10, anti-CBP, anti-RNA polymerase II, anti-p65, anti-IκBα, anti-P-CREB-Ser133 and anti-PKAc were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-P-p65-Ser536, anti-P-ERK-Thr202/Tyr204, anti-P-JNK-Thr183/Tyr185, anti-P-p38-Thr180/Tyr182, anti-P-MSK-1-Thr581, anti-Lamin A/C and anti-CREB were from Cell Signaling Technology (Danvers, MA, USA). Anti-Ac-H3-Lys27 and GAPDH were from AbCam (Cambridge, UK). Anti-α-tubulin and anti-α-actin were from Sigma-Aldrich. In figures, the expression “antibody anti-” was substituted by the Greek letter “α-”. AatII and HincII restriction enzymes were obtained from New England BioLabs (Ipswich, MA, USA).

Kolmus K., Van Troys M., Van Wesemael K., Ampe C., Haegeman G., Tavernier J, & Gerlo S. (2014). β-Agonists Selectively Modulate Proinflammatory Gene Expression in Skeletal Muscle Cells via Non-Canonical Nuclear Crosstalk Mechanisms. PLoS ONE, 9(3), e90649.

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Western Blot Analysis of Adrenergic Receptors in Rat Renal Cortex

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Total protein was extracted after homogenizing the rat renal cortex, and the concentration was determined with a bicinchoninic acid protein assay kit. The proteins were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then transferred to nitrocellulose membranes, which were incubated with the primary antibodies rabbit anti-β₁-AR (1:200), anti-β₂-AR (1:200), anti-β₃-AR (1:200), anti-α_1A-AR (1:200), anti-α_1B-AR (1:200), anti-α_1D-AR (1:200), anti-AT₁R (1:200), anti-AT₂R (1:200), and anti-GAPDH (1:200) (all from Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4 °C overnight, followed by incubation with goat anti-rabbit fluorescent (IRDye-conjugated) secondary antibodies (1:10,000; Rockland Immunochemicals, Gilbertsville, PA, USA) for 2 h at room temperature. The images were quantified by the Odyssey infrared imaging system (LI-COR Biosciences, Lincoln, NE, USA). Levels of proteins were normalized to that of GAPDH.

Peng Y., Li Y., Chen M., Song J., Jiang Z, & Shi S. (2020). High-dose nitrate therapy recovers the expression of subtypes α1 and β-adrenoceptors and Ang II receptors of the renal cortex in rats with myocardial infarction-induced heart failures. BMC Cardiovascular Disorders, 20, 99.

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Co-immunoprecipitation of β2-adrenergic receptor

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HEK293 cells stably expressing FLAG-tagged β₂-ARs50 (link) were transfected with Myc-tagged pIRES-EGFP-Myc-7TM or pIRES-EGFP-Myc-6TM constructs, or pIRES-EGFP-Myc empty vector (EV), and harvested at 48 h post-transfection. Pelleted cells were resuspended by passing through 22 g needle in ice-cold Pierce IP lysis buffer (Thermo Fisher Scientific). All further procedures were performed at +4 °C. After 15 min extraction, insoluble materials were removed by centrifugation (15,000 × g, 15 min) and the supernatant was used for IP. 400 μg of cellular extract was incubated with anti-FLAG M2 magnetic beads (Sigma-Aldrich) overnight. Beads were collected and washed 3 times with IP buffer and bound proteins were eluted with 3X FLAG peptide (Sigma-Aldrich), according to manufacturer’s instructions and then boiled in SDS-PAGE buffer. SDS-PAGE analysis and Western blotting were performed using the primary anti-β₂-AR (1:2,000; Santa Cruz Biotechnology) and anti-Myc (EQKLISEEDL) antibodies (1:1,500; EMD Millipore).

Samoshkin A., Convertino M., Viet C.T., Wieskopf J.S., Kambur O., Marcovitz J., Patel P., Stone L.S., Kalso E., Mogil J.S., Schmidt B.L., Maixner W., Dokholyan N.V, & Diatchenko L. (2015). Structural and functional interactions between six-transmembrane μ-opioid receptors and β2-adrenoreceptors modulate opioid signaling. Scientific Reports, 5, 18198.

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Immunofluorescence of DRG Neurons

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Fourteen micrometer (14 μm) frozen sections of DRGs (T13–L2) were used in the immunofluorescence study as described previously [31 (link), 32 (link)]. The primary antibodies were anti-P2X3R (1:100, Novus Biologicals, Minneapolis, USA), anti-β₂-AR (1:50, Santa Cruz Biotechnology, Texas, USA), gliocyte marker glutamine synthase (1:200, Abcam, Cambridge, UK), neuron marker NeuN (1:50, MAB377, Merck Millipore, Munich, Germany), large neuron marker NF200 (1:200, Abcam), small and medium peptidergic neuron marker CGRP (1:200, Abcam), small and medium non-peptidergic neuron marker IB4⁺ (1:500, Sigma, Munich, Germany). The secondary antibodies were Alexa Fluor 488 (1:100, Life Technologies Inc., Waltham, MA USA) and 555 (1:500, Life Technologies Inc.). Negative controls were without the primary antibodies.

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Antibody Validation for Immunoprecipitation and Blotting

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Antibodies employed in immunoprecipitation, Western blotting and immunostaining included: monoclonal M2 anti-FLAG
(Sigma-Aldrich Aldrich), anti-HA (Covance), anti-β₂AR (Santa Cruz), anti-nNOS (rabbit polyclonal (Santa
Cruz) or mouse monoclonal anti-nNOS (BD Bioscience)), anti-iNOS (rabbit polyclonal (Santa Cruz) or mouse monoclonal (BD
Bioscience)), anti-eNOS (rabbit polyclonal (Santa Cruz) or mouse monoclonal (BD Bioscience)), anti-ERK and anti-phospho ERK
(phospho-Thr202/Tyr204) (Cell Signaling), anti-PKD and anti-phospho-PDK (phospho-Ser916) (Cell Signaling), anti-MDM2 (Santa
Cruz), anti-p53 (Santa Cruz), anti-clathrin (heavy chain) (BD Transduction), anti-AP2 (BD Transduction), anti-β-actin
(Sigma-Aldrich) and anti-GAPDH (Millipore), anti-HIF-1α (R&D Systems), anti-pVHL (BD Bioscience).
Anti-βarr (A1CT and A2CT) rabbit polyclonal antibodies were provided by R.J. Lefkowitz, and additional antibodies
against βarr1/2 were obtained from Santa Cruz, Cell Signaling and BD Transduction. Validation of the specificity of
βarr1/2 antibodies in the setting of Western blotting is presented in Figures S1D and S1E.

Hayashi H., Hess D.T., Zhang R., Sugi K., Gao H., Tan B.L., Bowles D.E., Milano C.A., Jain M.K., Koch W.J, & Stamler J.S. (2018). S-nitrosylation of β-arrestins biases receptor signaling and confers ligand independence. Molecular cell, 70(3), 473-487.e6.

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