Sc 53646

Manufactured by Santa Cruz Biotechnology

Sc-53646 is a laboratory equipment product offered by Santa Cruz Biotechnology. It is designed for use in scientific research and analysis. The core function of this product is to facilitate the execution of specific experimental protocols and procedures within a laboratory setting.

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Lab products found in correlation

Bca kit, by Thermo Fisher Scientific (3 mentions) Ab6556, by Abcam (1 mentions) Supersignal west dura extended duration substrate, by Thermo Fisher Scientific (1 mentions) Cell lysis buffer, by Beyotime (1 mentions) Af1935, by R&D Systems (1 mentions) Iblot, by Thermo Fisher Scientific (1 mentions) Novex tris glycine sds sample buffer, by Thermo Fisher Scientific (1 mentions) Amersham ecl prime western blotting detection reagent, by GE Healthcare (1 mentions) Ta50011, by OriGene (1 mentions) Dc protein kit, by Bio-Rad (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Protease inhibitor, by Roche (1 mentions) Chemidoc, by Bio-Rad (1 mentions) Enhanced chemi luminescence (ecl), by PerkinElmer (1 mentions)

Spelling variants of this product

Anti‐α tubulin (10D8) (sc‐53646) (2 citations)

3 protocols using sc 53646

Validation of Anti-cPDSS2 Antibody

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An anti-cPDSS2 monoclonal antibody was custom made in mouse using the immunizing peptide IGISTWKEQV-amide corresponding to amino acid residues 221–230 (Abmart, Shanghai, China).
Cells were collected at 48 h after transfection. Protein was extracted from cells with Cell Lysis Buffer (Beyotime). The proteins (30 µg of total cell protein per lane) were separated by 12% SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes according to standard protocols. The membranes were blocked and incubated with anti-cPDSS2 (1∶1000), anti-GFP (1∶1000, ab6556, Abcam) and anti-alpha tubulin (1∶1000, sc-53646, Santa Cruz) antibodies overnight at 4°C. The Membranes were subsequently incubated with horseradish peroxidase (HRP) conjugated goat anti-mouse or goat anti-rabbit secondary antibodies (1∶10000) and visualized using SuperSignal West Dura Extended Duration Substrate (Thermo Scientific).

Feng C., Gao Y., Dorshorst B., Song C., Gu X., Li Q., Li J., Liu T., Rubin C.J., Zhao Y., Wang Y., Fei J., Li H., Chen K., Qu H., Shu D., Ashwell C., Da Y., Andersson L., Hu X, & Li N. (2014). A cis-Regulatory Mutation of PDSS2 Causes Silky-Feather in Chickens. PLoS Genetics, 10(8), e1004576.

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Western Blot Analysis of HIF1α

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Whole cell lysates were harvested in Novex® Tris-Glycine SDS Sample Buffer (ThermoFisher Scientific) then sonicated and incubated at 95°C for 5 mins. Samples were separated on a 10% Tris-Glycine gel then transferred onto PVDF membrane via iBlot (ThermoFisher Scientific). Membranes were blocked for 1 hr in 5% non-fat milk, TBST followed by overnight incubation at 4°C in primary HIF1α antibody (AF1935, R&D Systems) or monoclonal α-Tubulin antibody (SC-53646, Santa Cruz Biotechnology). Membranes were then washed in TBST and incubated with the appropriate secondary HRP-conjugated antibody (anti-goat for HIF1α, anti-mouse for α-Tubulin; Jackson ImmunoResearch Laboratories). Protein band visualization was performed using Amersham ECL Prime Western Blotting Detection Reagent (RPN2232, GE Healthcare).

Loftus S.K., Baxter L.L., Cronin J.C., Fufa T.D, & Pavan W.J. (2017). Hypoxia-induced HIF1α targets in melanocytes reveal a molecular profile associated with poor melanoma prognosis. Pigment cell & melanoma research, 30(3), 339-352.

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Western Blot Analysis of RNA Polymerase Subunits

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Cells were lysed in RIPA lysis buffer (50 mM Tris-HCl [pH 7.5], 150 mM NaCl, 1% NP-40, 0.1% SDS, and 1% sodium deoxycholate) supplemented with protease inhibitors (Roche), sonicated, and centrifuged at 13,200 rpm for 15 minutes. Protein concentrations were measured using Dc-Protein Kit (Bio-Rad) or BCA Kit (Thermo Fisher Scientific) and normalized against buffer only. Equal amounts of protein were separated on SDS-PAGE, transferred to PVDF membranes (Millipore Sigma), probed for target proteins, and detected using ECL (Perkin Elmer). The primary antibodies used for detection were RPA194 (C-1 1:500; Santa Cruz Biotechnology), RPA135 (goat polycloncal H-15 1:200; sc-17914 Santa Cruz Biotechnology), ZNRD1 (RPA12) (mouse monoclonal D10 1:200; Santa Cruz Biotechnology), DDK (mouse monoclonal TA50011 1:1000; Origene), α-tubulin (mouse monoclonal 10D8 1:1000; sc-53646 Santa Cruz Biotechnology), and GAPDH (rabbit monoclonal 14C10 1:5000; mAb#2118 Cell Signaling Technology). Horseradish peroxidase (HRP)-conjugated secondary antibodies were from DAKO or Santa Cruz Biotechnology. Chemiluminescence images were acquired using Bio-Rad ChemiDoc. Protein densitometry analysis was conducted using ImageJ software and normalized against loading control.

Ford B.L., Wei T., Liu H., Scull C.E., Najmi S.M., Pitts S., Fan W., Schneider D.A, & Laiho M. (2023). Expression of RNA polymerase I catalytic core is influenced by RPA12. PLOS ONE, 18(5), e0285660.

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