Srebp 1 2a4

Manufactured by Santa Cruz Biotechnology

Sourced in United States

SREBP-1 (2A4) is a mouse monoclonal antibody that recognizes the SREBP-1 protein. SREBP-1 is a transcription factor that regulates the expression of genes involved in lipid and cholesterol biosynthesis.

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Lab products found in correlation

Cycloheximide (chx), by Cell Signaling Technology (1 mentions) 9 cis retinoic acid, by Merck Group (1 mentions) Methyl β cyclodextrin, by Merck Group (1 mentions) Ly294002, by Cell Signaling Technology (1 mentions) C13 glucose, by Cambridge Isotopes (1 mentions) Gsk 2033, by Axon Medchem (1 mentions) Pparα, by Santa Cruz Biotechnology (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Ampkα1 2, by Santa Cruz Biotechnology (1 mentions) Hif 1α, by Santa Cruz Biotechnology (1 mentions) Aurora b, by Abcam (1 mentions) Srebp 2 1c6, by Santa Cruz Biotechnology (1 mentions) Aurora a, by Thermo Fisher Scientific (1 mentions) β actin, by Proteintech (1 mentions) Protease and phosphatase inhibitor cocktail, by Roche (1 mentions) P0013, by Beyotime (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions)

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Monoclonal anti-SREBP-1 (2A4), (2 citations)

4 protocols using srebp 1 2a4

Regulation of Lipid Metabolism Pathways

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Reagents were purchased from the following sources. Antibodies used are as follows: SREBP1 (2A4, Santa Cruz Biotechnology), SREBP2 (30682, Abcam), LXRα (PP-PPZ0412-00, R&D systems), LXRβ (K8917, R&D systems), phospho-T308 AKT (C31E5E), AKT (11E7) (Cell Signaling Technology).
Drugs used are as follows: Cycloheximide (Cell Signaling Technology) was used at 10 μg/ml. 9-cis-retinoic acid (Sigma) was used at 50 μM. The LXR antagonist GSK-2033 (Axon Medchem) was used at 500 nM. The LXR agonist GW3965 (Fisher Scientific) was used at 500 nM. PI3K inhibitor, LY294002 (Cell Signaling Technology), was used at 10 μM. Rapamycin was used at 100 nM and received as a gift from David Sabatini. Methyl-beta-cyclodextrin was purchased from Sigma. All sterols except for custom-synthesized ent-4β-HC (see below) were purchased from Steraloids. C13 glucose was purchased from Cambridge Isotope Laboratories.

Moldavski O., Zushin P.J., Berdan C.A., Van Eijkeren R.J., Jiang X., Qian M., Ory D.S., Covey D.F., Nomura D.K., Stahl A., Weiss E.J, & Zoncu R. (2021). 4β-Hydroxycholesterol is a prolipogenic factor that promotes SREBP1c expression and activity through the liver X receptor. Journal of Lipid Research, 62, 100051.

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Liver Protein Expression Analysis

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According to the methods of our previous study, total proteins were isolated from 50 mg of liver tissue and western blotting was performed. The blots were probed using the following antibodies: HIF-1α (sc-10790, 1:1000), PPARα (sc-9000, 1:1000), SREBP-1 (2A4) (sc-13551, 1:500), DEC1 (s-8) (sc-101023, 1:1000), AMPKα1/2 (sc-74461, 1:1000), Thr172-p-AMPKα1/2 (sc-33524, 1:1000), and β-actin (sc-477778, 1:1000); the above-mentioned antibodies were all from Santa Cruz Biotechnology, Dallas, TX, USA. ACC (#3662, Cell Signaling Technology, Inc., Danvers, MA, USA) and Ser79-p-ACC (#3661; Cell Signaling Technology, Inc., Danvers, MA, USA). The density of protein bands was analyzed using Bio-Rad imaging software (Bio-Rad Laboratories, Hercules, CA, USA). The individual values were originally expressed as a ratio of a standard (β-actin content) and then expressed as a fold change of the control group value.

Wang Y., Lavier J., Hua W., Gong L., Wei H., Wang J., Pellegrin M., Millet G.P, & Zhang Y. (2022). Effects of Six Weeks of Hypoxia Exposure on Hepatic Fatty Acid Metabolism in ApoE Knockout Mice Fed a High-Fat Diet. Life, 12(10), 1535.

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Molecular Signaling Pathways in Cancer

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TAK901 (#T2709) was purchased from TargetMol (Shanghai, China). Primary antibodies were as follows: SREBP-1 (2A4) (#sc-13551, Santa Cruz Biotechnology, Santa Cruz, CA, USA), SREBP-2 (1C6) (#sc-13552, Santa Cruz Biotechnology, Santa Cruz, CA, USA), phospho-GSK-3β (Ser9) (D85E12) (#5558, CST, Danvers, MA, USA), β-actin (#20536-1-AP, Proteintech, Wuhan, China), Ki67 (Servicebio, Wuhan, China),histone H3 (#4499s, CST, Danvers, MA, USA), phospho-histone H3 (Ser10) (#53348T, CST, Danvers, MA, USA), Aurora A (#45-8900, Invitrogen, Carlsbad, CA, USA), Aurora B (#ab2254, abcam, Waltham, MA, USA) and c-Myc (#9402S, CST, Danvers, MA, USA). Secondary antibodies were as follows: anti-mouse IgG, HRP-linked antibody (#7076, CST, Danvers, MA, USA), anti-rabbit IgG, HRP-linked antibody (#7074, CST, Danvers, MA, USA).

Zhan X., Qiu R., He Y., Zhao Z., Huang M., Liu Q., Zhi F, & Long W. (2022). The Aurora Kinase Inhibitor TAK901 Inhibits Glioblastoma Growth by Blocking SREBP1-Mediated Lipid Metabolism. Cancers, 14(23), 5805.

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Profiling Cellular Signaling Pathways with Western Blotting

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Tissue and cell lysates prepared in RIPA buffer (1% Triton X-100, 1% deoxycholate, 0.1% SDS, 10 mM Tris and 150 mM NaCl) with protease and phosphatase inhibitor cocktail (Roche, USA) were separated with SDS-PAGE and probed with primary antibodies against P-ERK (20G11), ERK (9102), P-FAK (3283), FAK (3285), P-AKT Ser473 (D9E), Akt (C67E7), Acc1 (C83B10), and Fasn (C20G5) from CST and Srebp1 (2A4) from Santa Cruz. After incubation with primary antibodies at 4 °C overnight, membranes were incubated with fluorescent secondary antibodies (goat anti-rabbit IRDye 800CW, goat anti-mouse IRDye 800CW, Licor Odyssey, United States). Membranes were examined by an Odyssey infrared imaging system (LI-COR Biosciences, United States). For coimmunoprecipitation of Sema7A and Itgb1, ADSCs were incubated with 10 μg/mL Sema7A for 3 h and lysed with lysis buffer (P0013, Beyotime Biotechnology, China). The lysates were precleared with protein G and immunoprecipitated with Sema7A antibody (18070-1-AP, Proteintech) or IgG. The coimmunoprecipitated proteins and input sample were blotted with Itgb1 antibody (sc-374429, A4, Santa Cruz).

Lu Q., Liu Z., Zhao L., Xu L., Liu C., Li L., Cao Y., Li F., Wu L., Wang L., Chen T., You T., Ren L., Wang G., Tang C, & Zhu L. (2023). Sema7A protects against high-fat diet-induced obesity and hepatic steatosis by regulating adipo/lipogenesis. Molecular Metabolism, 70, 101698.

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