Anti caveolin 3

Manufactured by BD

Sourced in United States

Anti-caveolin-3 is a laboratory reagent used in research applications. It is a protein that binds to and recognizes the caveolin-3 protein, which is involved in the formation of caveolae, small invaginations of the cell membrane. This reagent can be used to detect and study the expression and localization of caveolin-3 in biological samples.

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Lab products found in correlation

7 protocols using anti caveolin 3

Membrane Fraction Isolation and Western Blotting of β-Adrenergic Receptors

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Hearts were frozen in situ as above and a membrane fraction was isolated as previously described (Zhou et al., 1998 (link)), with modifications. Hearts were homogenized in hypotonic lysis buffer and the
homogenate was centrifuged at 2,000 G for 10 min. The supernatant was collected and centrifuged at 9,000 G for 20 min, and the
resultant supernatant was centrifuged at 180,000 G for 90 min. Essentially all β₁AR and
β₂AR were recovered in this final pellet. The pellet was reconstituted with RIPA buffer containing
protease inhibitor (Roche) then running buffer for Western blotting (anti-β₁AR (Santa Cruz),
anti-β₂AR (as above) and anti-Caveolin3 (BD Biosciences)).

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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Skeletal Muscle Cell Culture Protocol

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Dulbecco’s modified Eagle’s medium, Minimal Essential Medium Eagle alpha modification (α-MEM), neomycin, fetal bovine serum, horse serum, and penicillin-streptomycin were obtained from Thermo Fischer Scientific (Waltham, MA, USA). Collagenase type II was from Worthington Biochemical (Lakewood, NJ, USA). Complete™ Mini protease inhibitors were from Roche Applied Science (Indianapolis, IN, USA). Protein A/G agarose was from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse anti-Strep antibody was from IBA GmbH (Gottingen, Germany), anti-caveolin-3 was from BD Bioscience (San José, CA, USA), and anti-Dys was from Leica Biosystems (Wetzlar, Germany). Rabbit anti-P2Y₂R and anti-Panx1 antibodies, mouse anti-DHPR α-1 antibody, and secondary anti-mouse and anti-rabbit antibodies conjugated to either horseradish peroxidase or Alexa Fluor® dyes were from Thermo Fischer Scientific. Enhanced chemiluminescence (ECL) reagents were from Amersham Biosciences (Piscataway, NJ, USA). ECM (gel from Engelbreth-Holm-Swarm murine sarcoma) matrix was from Sigma-Aldrich (St. Louis, MO, USA). All other reagents used were of analytical quality.

Arias-Calderón M., Almarza G., Díaz-Vegas A., Contreras-Ferrat A., Valladares D., Casas M., Toledo H., Jaimovich E, & Buvinic S. (2016). Characterization of a multiprotein complex involved in excitation-transcription coupling of skeletal muscle. Skeletal Muscle, 6, 15.

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Antibody Sources for Cellular Imaging

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Different anti-OXPHOS antibodies were obtained from Molecular Probes (Carlsbad,
California, USA) and Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Anti-cytochrome c, anti-caveolin-1 and anti-caveolin-3
antibodies were obtained from BD Biosciences (Franklin Lakes, New Jersey, USA).
MitoTracker Red FM dye and Alexa Fluor 488-conjugated anti-mouse IgG were
purchased from Invitrogen (Carlsbad, California, USA). FITC-conjugated
anti-mouse IgG and Rhodamine-conjugated anti-rabbit IgG were obtained from Abcam
(Cambridge, UK). Collagenase type I and bovine serum albumin (BSA) were
purchased from Sigma (St. Louis, Missouri, USA).

Lee H., Kim S.H., Lee J.S., Yang Y.H., Nam J.M., Kim B.W, & Ko Y.G. (2016). Mitochondrial oxidative phosphorylation complexes exist in the sarcolemma of skeletal muscle. BMB Reports, 49(2), 116-121.

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Antibody Reagents for Cell Signaling

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Unless indicated otherwise, all reagents were obtained from Sigma Chemical Company (Gillingham, UK) and were of the highest grade available. Anti-clathrin heavy chain and anti-caveolin 3 were from BD Biosciences (Oxford, UK). Anti-HADHA, cavin1, GFP, and Glut4 antibodies were from Abcam (Cambridge, UK). Anti-sodium pump α2 subunit and integrin α5 were from Merck Millipore (Watford, UK). Anti-adenylyl cyclase 5/6, insulin receptor, and PKCε were from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-Giα1/2 was from Pierce Antibodies (Thermo Scientific, Rockford, IL). Anti-moesin and anti-Kir 6.2 were from the MRC Protein Phosphorylation Unit, University of Dundee. The monoclonal antibody α6F raised against the sodium pump α1 subunit by Douglas M. Fambrough were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Department of Biology, Iowa City, IA 52242. An antibody to PLM is described elsewhere (40 (link)). Protease and phosphatase inhibitor cocktails were from Merck Millipore and Sigma, respectively.

Wypijewski K.J., Tinti M., Chen W., Lamont D., Ashford M.L., Calaghan S.C, & Fuller W. (2015). Identification of Caveolar Resident Proteins in Ventricular Myocytes Using a Quantitative Proteomic Approach: Dynamic Changes in Caveolar Composition Following Adrenoceptor Activation. Molecular & Cellular Proteomics : MCP, 14(3), 596-608.

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Sodium Pump α1 Subunit Antibody Protocol

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Anti-NCX1 was from Swant, anti-caveolin 3 and flotillin 2 were from BD Biosciences (San Jose, CA, USA), and anti–green fluorescent protein was from Abcam (Cambridge, MA, USA). The monoclonal antibody α6F raised against the sodium pump α1 subunit by Douglas M. Fambrough (Johns Hopkins University, Baltimore, MD, USA) was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD) and maintained by The University of Iowa, Department of Biology (Iowa City, IA, USA).

Reilly L., Howie J., Wypijewski K., Ashford M.L., Hilgemann D.W, & Fuller W. (2015). Palmitoylation of the Na/Ca exchanger cytoplasmic loop controls its inactivation and internalization during stress signaling. The FASEB Journal, 29(11), 4532-4543.

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Immunofluorescence Staining of Paraformaldehyde-Fixed Cells

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Cells fixed in 4% paraformaldehyde were treated with 0.15% Triton X-100 and stained according to previously published protocols [15 (link)]. The following primary antibodies were used: anti-Samp1 (from Hallberg laboratory); anti-lamin B and anti-pericentrin from Abcam (Cambridge, UK); anti-desmin and anti-emerin from Monosan (Uden, The Netherlands); anti-SUN1 and anti-SUN2 from Sigma (St. Louis, MO, USA); anti-farnesylated prelamin A (1188-2) from Diatheva (Pesaro, Italy), anti-caveolin 3 from BD Transduction (San Francisco, CA, USA); anti-prelamin A (Sc-6214) form Santa Cruz Biotechnology (Dallas, TX, USA), and anti-laminin alpha2 from Chemicon (Massachusetts, MA, USA). Image acquisition was performed by using a Nikon Eclipse Ni epifluorescence microscope equipped with a digital CCD camera and NIS-Elements 4.3 AR software. Photoshop CS was used for image processing. Mean fluorescence intensity was measured by using NIS-Elements 4.3 AR.

Mattioli E., Columbaro M., Jafferali M.H., Schena E., Hallberg E, & Lattanzi G. (2018). Samp1 Mislocalization in Emery-Dreifuss Muscular Dystrophy. Cells, 7(10), 170.

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Immunofluorescence Staining of Cryostat Sections

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Immunofluorescence staining was performed on 8 μm-thick cryostat sections using the following antibodies: anti-phospholamban-PLN (1:200, rabbit polyclonal; Invitrogen Life Technologies, Carlsbad, CA, USA), anti-MYH1 (1:100, mouse monoclonal; Abcam, Cambridge, UK), anti-p62 (1:100, rabbit polyclonal; Abcam), anti-LC3 (1:100, rabbit polyclonal; ThermoFisher, Waltham, MA, USA), anti-valosin-containing protein VCP (1:100 mouse monoclonal; ThermoFisher) and anti-caveolin-3 (1:1000, mouse monoclonal; BD Transduction, East Rutherford, NJ, USA). Slides were then incubated with the appropriate secondary antibody conjugated to Alexa 488 or Alexa 568 (1:2000, Invitrogen Life Technologies). Sections were mounted in Vectashield anti-fade mounting medium containing DAPI (Vector Laboratories, Burlingame, CA, USA). Image fields were acquired using optical microscope Leica DM4000B equipped with camera (DFC420C).

Zanotti S., Ripolone M., Napoli L., Velardo D., Salani S., Ciscato P., Priori S., Kukavica D., Mazzanti A., Diamanti L., Vegezzi E., Moggio M., Corti S., Comi G, & Sciacco M. (2023). Characterization of Skeletal Muscle Biopsy and Derived Myoblasts in a Patient Carrying Arg14del Mutation in Phospholamban Gene. Cells, 12(10), 1405.

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