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Sc10646

Manufactured by Santa Cruz Biotechnology
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SC10646 is a laboratory equipment product manufactured by Santa Cruz Biotechnology. It serves as a centrifuge, a device used to separate components of a liquid mixture based on their relative densities by applying centrifugal force.

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Lab products found in correlation

Ab2912, by Abcam (2 mentions) Ab11425, by Abcam (2 mentions) Rabbit igg sc 2027, by Santa Cruz Biotechnology (2 mentions) Dynabeads protein g, by Thermo Fisher Scientific (2 mentions) Quick start bradford dye reagent, by Bio-Rad (2 mentions) Bis tris gel, by Bio-Rad (2 mentions) Mouse igg sc 2025, by Santa Cruz Biotechnology (2 mentions) Dynabeads protein g, by Santa Cruz Biotechnology (2 mentions) Ac003, by Alomone (2 mentions) Ac062, by Alomone (2 mentions) Goat igg sc2028, by Santa Cruz Biotechnology (2 mentions) Ab59791, by Abcam (1 mentions) Sc 21713, by Santa Cruz Biotechnology (1 mentions) Dm2500, by Leica Microsystems (1 mentions) Ab19031, by Abcam (1 mentions) Protease and phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions) Ab50533, by Abcam (1 mentions) Ab3612, by Abcam (1 mentions) Donkey anti rabbit igg antibodies, by LI COR (1 mentions) Sc 48166, by Santa Cruz Biotechnology (1 mentions)

4 protocols using sc10646

1

Cardiac Protein Immunoprecipitation and Analysis

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The study was carried out following the Guide for the Care and Use of Laboratory Animals published by the United States National Institutes of Health and the Directive 2010/63/EU of the European Parliament. 6-8 weeks old male mice of strain C57BL6 were sacrificed by cervical dislocation and their hearts were harvested and snap frozen in liquid nitrogen and stored at -80 °C. Heart tissue was homogenized on a Precellys 24 and solubilized in ice-cold lysis buffer containing protease and phosphatase inhibitors. Tissue lysates were centrifuged to remove insoluble debris. For each tissue preparation produced, lysates derived from 5 mice were pooled and protein concentrations were measured by Quick Start Bradford Dye Reagent (Biorad). Solubilized heart tissue lysate was pre-cleared with Dynabeads protein G (Invitrogen) before incubation with primary antibody followed by binding to Dynabeads protein G, using either anti-KCNQ1 (10 μl SC10646, Santa Cruz), anti-CACNA1C (2 μl AC003, Alomone), anti-KCNH2 (2 μl AC062, Alomone), anti-CAV3 (2 μl ab2912, Abcam), anti-SNTA1 (2 μl ab11425, Abcam) or control IgG (1.5 μl goat IgG: SC2028, 1.5 μl rabbit IgG: SC2027, 1.5 μl mouse IgG: SC2025, Santa Cruz). After washing, bound proteins were eluted with 1× sample buffer containing 100 mM dithiothreitol (70 °C, 3 min) and separated by SDS-PAGE (4-15 % Bis-Tris gels, BioRad).
Lundby A., Rossin E.J., Steffensen A.B., Rav Acha M., Newton-Cheh C., Pfeufer A., Lynch S.N., Olesen S.P., Brunak S., Ellinor P.T., Jukema J.W., Trompet S., Ford I., Macfarlane P.W., Krijthe B.P., Hofman A., Uitterlinden A.G., Stricker B.H., Nathoe H.M., Spiering W., Daly M.J., Asselbergs F.W., van der Harst P., Milan D.J., de Bakker P.I., Lage K, & Olsen J.V. (2014). Annotation of loci from genome-wide association studies using tissue-specific quantitative interaction proteomics. Nature methods, 11(8), 868-874.
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2

Cardiac Protein Immunoprecipitation and Analysis

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The study was carried out following the Guide for the Care and Use of Laboratory Animals published by the United States National Institutes of Health and the Directive 2010/63/EU of the European Parliament. 6-8 weeks old male mice of strain C57BL6 were sacrificed by cervical dislocation and their hearts were harvested and snap frozen in liquid nitrogen and stored at -80 °C. Heart tissue was homogenized on a Precellys 24 and solubilized in ice-cold lysis buffer containing protease and phosphatase inhibitors. Tissue lysates were centrifuged to remove insoluble debris. For each tissue preparation produced, lysates derived from 5 mice were pooled and protein concentrations were measured by Quick Start Bradford Dye Reagent (Biorad). Solubilized heart tissue lysate was pre-cleared with Dynabeads protein G (Invitrogen) before incubation with primary antibody followed by binding to Dynabeads protein G, using either anti-KCNQ1 (10 μl SC10646, Santa Cruz), anti-CACNA1C (2 μl AC003, Alomone), anti-KCNH2 (2 μl AC062, Alomone), anti-CAV3 (2 μl ab2912, Abcam), anti-SNTA1 (2 μl ab11425, Abcam) or control IgG (1.5 μl goat IgG: SC2028, 1.5 μl rabbit IgG: SC2027, 1.5 μl mouse IgG: SC2025, Santa Cruz). After washing, bound proteins were eluted with 1× sample buffer containing 100 mM dithiothreitol (70 °C, 3 min) and separated by SDS-PAGE (4-15 % Bis-Tris gels, BioRad).
Lundby A., Rossin E.J., Steffensen A.B., Rav Acha M., Newton-Cheh C., Pfeufer A., Lynch S.N., Olesen S.P., Brunak S., Ellinor P.T., Jukema J.W., Trompet S., Ford I., Macfarlane P.W., Krijthe B.P., Hofman A., Uitterlinden A.G., Stricker B.H., Nathoe H.M., Spiering W., Daly M.J., Asselbergs F.W., van der Harst P., Milan D.J., de Bakker P.I., Lage K, & Olsen J.V. (2014). Annotation of loci from genome-wide association studies using tissue-specific quantitative interaction proteomics. Nature methods, 11(8), 868-874.
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3

Immunolabeling of Cochlear Markers

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Cochleae were dissected from saline-perfused, formalin-fixed temporal bones of young adult female M. fascicularis (3 years old, N = 3) and decalcified with 0.1 M EDTA/phosphate buffer at 4°C for at least 6 weeks, then embedded in paraffin. The 5 μm sections were rehydrated, pretreated with sodium citrate buffer, then blocked in PBS containing 5% FBS, 1% BSA, and 0.05% Tween-20. The primary antibodies used in this study were rabbit antiserum against SLC12A2, generated using the amino-terminal region of the protein as an immunogen, and thus able to detect both the exon 21-included and -skipped isoforms (Abcam: ab59791, 1:100), mouse monoclonal antibody against ATP1B1 (Santa Cruz Biotechnology: sc21713, 1:400), and goat antiserum against KCNQ1 (Santa Cruz Biotechnology: sc10646, 1:200). Sections were incubated with primary antibodies at 4°C overnight, and signals were visualized using appropriate secondary antibodies, conjugated with either Alexa 488 or 568, followed by counterstaining with DAPI. Images were captured by fluorescence (DM2500, Leica Microsystems) and confocal (Axio 700) microscopy.
Mutai H., Wasano K., Momozawa Y., Kamatani Y., Miya F., Masuda S., Morimoto N., Nara K., Takahashi S., Tsunoda T., Homma K., Kubo M, & Matsunaga T. (2020). Variants encoding a restricted carboxy-terminal domain of SLC12A2 cause hereditary hearing loss in humans. PLoS Genetics, 16(4), e1008643.
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4

KCNQ1, KCNE1, and α1A-AR Protein Expression

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Cells were transfected with GFP-tagged KCNQ1 (1.5 μg) and KCNE1 (1.5 μg), and α1A-AR (3 μg) (for phenylephrine experiments). Cells were harvested with Laemmli buffer supplemented with phosphatase and protease inhibitor (Thermo Fisher Scientific). Samples were sonicated, centrifuged for 2min at 15,000 g and supernatants were harvested as whole cell lysate. The samples were run on a 7% Acrylamide gel or with 4–20% Precast Protein Gels (Bio-Rad) under 95 V for 2h. Antibody against KCNQ1, or GAPDH (respectively sc-10646 and sc-48166, Santa Cruz technology) were used and recognized respectively by donkey anti-goat IgG or donkey anti-rabbit IgG antibodies (LI-COR Biosciences). For Rab5 experiments, antibody against Rab5 (C8B1, Rabbit mAb from Cell Signaling Technology) was used and recognized by donkey anti-rabbit IgG antibodies (LI-COR Biosciences). For Rab7 experiments, antibody against Rab7 (Rab7–117, mouse monoclonal from abcam, #ab50533) was used. For Rab11 experiments, antibody against Rab11 (Rabbit polyclonal from abcam, #ab3612) was used. cPKC isoforms were recognized by an antibody against cPKC (Rabbit polyclonal from abcam, ab19031). The membranes were visualized using LI-COR (LI-COR Biosciences) and analyzed with ImageJ software.
Parks X.X., Ronzier E., O-Uchi J, & Lopes C.M. (2019). Fluvastatin inhibits Rab5-mediated IKs internalization caused by chronic Ca2+-dependent PKC activation. Journal of molecular and cellular cardiology, 129, 314-325.
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