Ab10983

Manufactured by Santa Cruz Biotechnology

Ab10983 is a laboratory product manufactured by Santa Cruz Biotechnology. It is a purified antibody that can be used for various research applications. The core function of this product is to detect and bind to specific target molecules in samples. No further details about its intended use can be provided in an unbiased and factual manner.

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Lab products found in correlation

Anti β actin, by Merck Group (3 mentions) Ab118573, by Abcam (2 mentions) Ab55744, by Merck Group (2 mentions) Horseradish peroxidase, by GE Healthcare (2 mentions) Enhanced chemiluminescence detection system, by GE Healthcare (2 mentions) A1978, by Merck Group (2 mentions) Hybond nitrocellulose membrane, by GE Healthcare (2 mentions) Sc 18661, by Abcam (2 mentions) Chemi capt software, by Avantor (2 mentions) T6074, by Merck Group (2 mentions) F3165, by Merck Group (2 mentions) Protease inhibitor cocktail, by Roche (2 mentions) Oxphos cocktail, by Abcam (1 mentions) Sc 7947, by Merck Group (1 mentions) Sc 8035, by Abcam (1 mentions) Streptavidin hrp, by Thermo Fisher Scientific (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Bradford assay, by Bio-Rad (1 mentions) Hpa006111, by Abcam (1 mentions) Anti vdr, by Santa Cruz Biotechnology (1 mentions)

5 protocols using ab10983

Protein Extraction and Analysis from Mammalian Cells

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Homogenized tissue or cells were lysed with ice-cold RIPA buffer (Pierce, 89900) containing 1X protease inhibitor cocktail (GenDEPOT, P3100) and incubated at 4 °C for 30 min. After centrifugation at 15,000 × g for 15 min, the supernatant was moved to a new tube. Protein concentrations were measured using the Bradford assay (Bio-Rad, 5000001). Protein samples were directly analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with SDS sample buffer (60 mM Tris-Cl (pH 6.8), 10% sodium lauryl sulfate, 25% glycerol, 100 mM dithiothreitol, 0.04% Bromophenol blue) without boiling. Western blot analysis was performed according to standard methods. Antibodies used in immunoblot analyses included those against LETMD1 (LSBio, LS-C384640, 1:1000); UCP1 (Abcam, ab10983, 1:1000); HSP90 (Santa Cruz, sc-7947, 1:2000); α-Tubulin (Sigma-Aldrich, sc-8035, 1:3000); OXPHOS cocktail (Abcam, ab110413, 1:1000); Streptavidin-HRP (Thermo Scientific, S911, 1:3000); and Flag (Sigma-Aldrich, F3165, 1:3000). The specific signals were amplified by horseradish peroxidase-conjugated secondary anti-rabbit or anti-mouse antibody (Santa Cruz, 1:3000).

Park A., Kim K.E., Park I., Lee S.H., Park K.Y., Jung M., Li X., Sleiman M.B., Lee S.J., Kim D.S., Kim J., Lim D.S., Woo E.J., Lee E.W., Han B.S., Oh K.J., Lee S.C., Auwerx J., Mun J.Y., Rhee H.W., Kim W.K., Bae K.H, & Suh J.M. (2023). Mitochondrial matrix protein LETMD1 maintains thermogenic capacity of brown adipose tissue in male mice. Nature Communications, 14, 3746.

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Immunoblotting Analysis of Adipose Tissue

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Experiments were performed as described^{1 (link)}. Fat tissues were grounded in RIPA buffer [50 mM Tris pH 7.5, 1 % NP40, 0.5 % sodium deoxycholate, 0.1 % SDS, 150 mM NaCl, 5 mM EDTA, and protease inhibitor cocktail (45 μg/ml, Roche, 11 873 580 001)] at 4 °C. Homogenates were separated in 10 % polyacrylamide gels and blotted to Hybond nitrocellulose membranes (GE Healthcare). Membranes were decorated using following antibodies: anti-LSD1^{1 (link)} (1/1000), anti-Prdm16 (abcam, ab118573, 1/500), anti-Ucp1 (abcam, ab10983, 1/1000), Fabp4 (Santa-Cruz, sc-18661, 1/2000), anti-Nrf1 (abcam, ab55744, 1/1000), anti-Flag (Sigma, F3165, 1/500), anti-β-Tubulin (Sigma, T6074, 1/10000), or anti-β-Actin (Sigma, A1978, 1/10000). Secondary antibodies conjugated to horseradish peroxidase (GE Healthcare) were detected using an enhanced chemiluminescence detection system (GE Healthcare). Protein levels were quantified using the Chemi Capt software (Peqlab). Uncropped scans are available in Supplementary Fig. 16.

Duteil D., Metzger E., Willmann D., Karagianni P., Friedrichs N., Greschik H., Günther T., Buettner R., Talianidis I., Metzger D, & Schüle R. (2014). LSD1 promotes oxidative metabolism of white adipose tissue. Nature communications, 5, 4093.

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Immunoblotting Analysis of Adipose Tissue

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Immunoblot Analysis of Adipose Tissue Proteins

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Cells were washed with PBS and lysed in protein lysis buffer (1% Triton
X-100, 50 mM HEPES at pH 7.4, 150 mM NaCl, 1% glycerol, 2 mM EDTA,
protease/phosphatase inhibitor cocktail). For immunoblot analysis of surgically
dissected fat tissue depots, tissues were cryomilled and lysed in RIPA buffer
(150 mM NaCl, 50 mM HEPES at pH 7.4, 0.1% SDS, 1% Triton X-100, 1% glycerol, 2
mM EDTA, 0.5% deoxycholate) containing a protease and phosphatase inhibitor
cocktail. Protein lysates were mixed with 5X SDS sample buffer and boiled,
separated by SDS-PAGE, transferred to polyvinylidene difluoride (PVDF) membrane,
and subjected to immunoblot analysis. Immunoblot analysis was subsequently
performed using the indicated antibodies and indicated dilutions: 1:10,000 GS
(BD#610517), 1:1,000 UCP1 (CST#14670 for tissue and Abcam#ab10983 for cells),
1:5,000 VINC (SantaCruz#sc-73614), 1:5,000 GLS (ProteinTech#12855), 1:2,000
GLUD1 (ProtienTech#14299), 1:1,000 FASN (CST#3180), 1:1,000 PPARy
(CST#2443).

Kanangat S., Meduri G.U., Tolley E.A., Patterson D.R., Meduri C.U., Pak C., Griffin J.P., Bronze M.S, & Schaberg D.R. (1999). Effects of Cytokines and Endotoxin on the Intracellular Growth of Bacteria. Infection and Immunity, 67(6), 2834-2840.

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Western Blot Analysis of Protein Targets

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Tissues were lysed in radioimmunoprecipitation assay (RIPA) buffer containing 0.1% SDS, 1% NP-40, 0.5% Na deoxycholate, 150 mM NaCl, 50 mM Tris-Cl (pH 8.0), 1 mM EDTA supplemented with protease inhibitor (Roche). Proteins were separated by SDS-PAGE, transferred to nitrocellulose membrane, and detected with the antibodies anti-NFIA (1:1000 dilution, Sigma HPA006111), anti-UCP1 (1:2000 dilution, Abcam, ab10983), anti-VDR (1:100 dilution, Santa Cruz Biotechnology, sc-13133), anti-FLAG M2 (Sigma F3165), and anti-β actin (Sigma A3854).

Hiraike Y., Tsutsumi S., Wada T., Oguchi M., Saito K., Nakamura M., Ota S., Koebis M., Nakao H., Aiba A., Nagano G., Ohno H., Oki K., Yoneda M., Kadowaki T., Aburatani H., Waki H, & Yamauchi T. (2022). NFIA determines the cis-effect of genetic variation on Ucp1 expression in murine thermogenic adipocytes. iScience, 25(8), 104729.

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