C ebp

Manufactured by Santa Cruz Biotechnology

Sourced in United States

C/EBP is a family of transcription factors that play a role in the regulation of gene expression. These factors are involved in cellular processes such as cell differentiation, metabolism, and immune response. The C/EBP family includes several isoforms, each with distinct functions. This product provides a tool for researchers to study the role of C/EBP in various biological systems.

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5 protocols using c ebp

Transcription Factor Profiling

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Oligonucleotides used are given in Table 1. The prefix “r” denotes the ribonucleotide where appropriate. Antibodies against C/EBP (catalog SC-746, 1:1000), USF-2 (SC-862, 1:1000), E2A (N-649, 1:2000), MyoD (SC-760, 1:2000), and NFκB (SC-372, 1:1000) were from Santa Cruz Biotechnology (Dallas, TX, USA). Dilutions given are those used for Western blots. Antibody against c-Myc (catalog #9402, 1:1000) was from Cell Signaling Technology (Beverly, MA, USA). Ultralink^© Hydrazide Agarose was from Thermo Scientific (Rockford, IL, USA). All chemicals were of the highest purity available commercially.

Jia Y, & Jarrett H.W. (2015). Method for trapping affinity chromatography of transcription factors using aldehyde-hydrazide coupling to agarose. Analytical biochemistry, 482, 1-6.

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Rabbit Polyclonal Antibody Generation and Characterization

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The custom rabbit polyclonal antiserum against PARP-1 used for Western blotting and ChIP assays was generated by using a purified recombinant antigen comprising the amino-terminal half of PARP-1 (42 (link)) (now available from Active Motif; cat. no. 39559). The custom rabbit polyclonal antiserum against NMNAT-1 was raised against purified recombinant human and mouse NMNAT-1 (Pocono Rabbit Farm and Laboratory). The custom recombinant antibody-like anti-poly-ADP-ribose binding reagent (anti-PAR) was generated and purified in-house (now available from EMD Millipore, MABE1031). The other antibodies used were as follows: C/EBP (Santa Cruz, sc-150X), NMNAT-2 (Abcam, ab56980), -Tubulin (Abcam, ab6046), SIRT1 [custom rabbit polyclonal antiserum raised against mouse SIRT1 (35 (link))], acetyl-p53 K379 (Cell signaling, #2570), p53 (Cell signaling, #2524), H4K16Ac (Millipore, 07–329), Histone H4 (Millipore, 07–108), rabbit IgG (Invitrogen, 10500C), goat anti-rabbit HRP-conjugated IgG (Pierce, 31460), and goat anti-mouse HRP-conjugated IgG (Pierce, 31430).

Ryu K.W., Nandu T., Kim J., Challa S., DeBerardinis R.J, & Kraus W.L. (2018). Metabolic Regulation of Transcription Through Compartmentalized NAD+ Biosynthesis. Science (New York, N.Y.), 360(6389), eaan5780.

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Mapping Transcription Factor Binding in Vascular Cells

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Data from ENCODE^{26 (link)} were explored as described in eMethods. ChIP-seq experiments were performed on confluent HCASMC (Cell Applications 350-05a & Lonza CC-2583; cultured in SmGM-2 BulletKit media; Lonza) as described.^{27 (link)} TCF21 (Abcam ab49475), Jun (Santa Cruz Biotechnology sc-1694), JunD (Santa Cruz Biotechnology sc-74), and CEBP (Santa Cruz Biotechnology sc-150) transcription factor binding was interrogated and H3K27ac data were acquired using the same ChIP protocol with an anti-H3K27ac antibody (Abcam; ab4729). Reads were aligned to the human genome (GRCh37p13) using STAR.^{28 (link)}

Saleheen D., Zhao W., Young R., Nelson C.P., Ho W., Ferguson J.F., Rasheed A., Ou K., Nurnberg S.T., Bauer R.C., Goel A., Do R., Stewart A.F., Hartiala J., Zhang W., Thorleifsson G., Strawbridge R.J., Sinisalo J., Kanoni S., Sedaghat S., Marouli E., Kristiansson K., Zhao J.H., Scott R., Gauguier D., Shah S.H., Smith A.V., van Zuydam N., Cox A.J., Willenborg C., Kessler T., Zeng L., Province M.A., Ganna A., Lind L., Pedersen N.L., White C.C., Joensuu A., Kleber M.E., Hall A.S., März W., Salomaa V., O’Donnell C., Ingelsson E., Feitosa M.F., Erdmann J., Bowden D.W., Palmer C.N., Gudnason V., De Faire U., Zalloua P., Wareham N., Thompson J.R., Kuulasmaa K., Dedoussis G., Perola M., Dehghan A., Chambers J.C., Kooner J., Allayee H., Deloukas P., McPherson R., Stefansson K., Schunkert H., Kathiresan S., Farrall M., Frossard P.M., Rader D.J., Samani N.J, & Reilly M.P. (2017). Loss of Cardio-Protective Effects at the ADAMTS7 Locus Due to Gene-Smoking Interactions. Circulation, 135(24), 2336-2353.

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Comprehensive Immunological Assays Protocol

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The fluorochrome-labeled antibodies for flow cytometry were purchased from eBioscience (San Diego, CA). The antibodies of IL-10, A20, HDAC11, Sp, IRF, AP, CREB, c/EBP, c-Maf, NF-κB, STAT3, HDAC11 shRNA kit and A20 shRNA kit were purchased from Santa Cruz Biotech (Santa Cruz, CA). LPS, ChIP kit and luciferase kit were purchased from Sigma Aldrich (St. Louis., MO). The recombinant IL-4, IL-5, IL-13 and the ELISA kit of IL-4 were purchased from R&D Systems (Minneapolis, MN). The OVA-specific IgE ELISA kit was purchased from the Biomart (Beijing, China). The reagent kits for magnetic cell sorting were purchased from Miltenyi Biotech (San Diego, CA). Reagents for RT-qPCR and Western blotting were purchased from Invitrogen (Carlsbad, CA).

Li M.Y., Zhu M., Linghu E.Q., Feng F., Zhu B., Wu C, & Guo M.Z. (2016). Interleukin-13 suppresses interleukin-10 via inhibiting A20 in peripheral B cells of patients with food allergy. Oncotarget, 7(48), 79914-79924.

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Protein Extraction and Immunoblotting of Mouse Tissues

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Snap-frozen mouse tissues were ground in liquid nitrogen and lysed with lysis buffer (20 mM HEPES pH 7.5, 200 mM NaCl, 1 mM EDTA, 1 mM EGTA, 0.5% Triton X-100, 20 mM Na-pyrophosphate, 50 mM β-glycerophosphate and 50 mM NaF) containing a protease inhibitor cocktail (Tech and Innovation, Chuncheon, Korea). Antibodies against ATG7, p62, SDHA, PDH, LC3 (Cell Signaling Technology, Danvers, MA, USA), C/EBP (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and UCP1 (Abcam, Cambridge, UK) were used for immunoblotting analysis as indicated in the figures. Expression of HSP90 (Santa Cruz Biotechnology, Inc.) was measured as a gel loading control.

Kim D., Kim J.H., Kang Y.H., Kim J.S., Yun S.C., Kang S.W, & Song Y. (2019). Suppression of Brown Adipocyte Autophagy Improves Energy Metabolism by Regulating Mitochondrial Turnover. International Journal of Molecular Sciences, 20(14), 3520.

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