Sc 514451

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Sc-514451 is a laboratory reagent product offered by Santa Cruz Biotechnology. It is a chemical compound used in research applications. The core function of Sc-514451 is to serve as a research tool, without further interpretation of its intended use.

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Lab products found in correlation

4 protocols using sc 514451

Protein Expression Analysis Protocol

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Total cell extracts were prepared in 1× sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) sample loading buffer. Cell fractions were extracted with nuclear and cytoplasm protein extraction kit (Wanleibio, China). Cell proteins were resolved by SDS-PAGE, transferred to a polyvinylidene difluoride membrane, and probed with GAPDH (1:1000, BM3876, Bosterbio, USA), p-PERK, p-IRE1, ATF4, P16 (1:500, bs-3330R, bs-16698R, bs-1531R, bs-20656R, Bioss, China), H3, NRF2 (1:500, D153567, D121053, Sangon Biotech, China), and TNF-α, IL-6, P21, MT1, MT2, HRD1, VCP, P65, p-P65, IKK (1:200, sc-52746, sc-32296, sc-136020, sc-13180, sc-13177, sc-293484, sc-136273, sc-514451, sc-166748, sc-7606, Santa Cruz, USA). Secondary anti-rabbit, anti-mouse antibodies (1:1000, BM2004, BA1001, Bosterbio, USA) conjugated with horseradish peroxidase were used. Detection was performed using a Thermo Scientific Pierce enhanced chemiluminescence western blotting substrate (Thermo Scientific). Results were analyzed by Tanon-410 automatic gel imaging system (Shanghai Tianneng Corporation, China).

Fang J., Yan Y., Teng X., Wen X., Li N., Peng S., Liu W., Donadeu F.X., Zhao S, & Hua J. (2018). Melatonin prevents senescence of canine adipose-derived mesenchymal stem cells through activating NRF2 and inhibiting ER stress. Aging (Albany NY), 10(10), 2954-2972.

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MAP3K7 Signaling Pathway Analysis

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The cells were harvested in ice‐cold PBS, 48–72 h after transfection. Lysate samples were prepared by homogenizing the harvested HEK293Tcells in lysate buffer (10 mM Tris‐HCl 6.8, 2.5% SDS, 2 mM EDTA) containing protease inhibitor cocktail 2 (#P5726; Sigma) and 3 (#P0044; Sigma‐Aldrich) and protease inhibitor (#P8340; Sigma‐Aldrich). Protein concentrations were determined using the BCA kit (Pierce). Final working protein concentrations were adjusted to 1 mg/ml. Western blots were probed with primary antibodies against MAP3K7 (sc‐7967, 1:1000; Santa Cruz), phospho‐MAP3K7 (Thr187; #4536, 1:1000; Cell Signaling), extracellular signal‐regulated kinase (ERK)1/2 (#9102, 1:2000; Cell Signaling), phospho‐ERK1/2 (#9101, 1:2000; Cell Signaling), actin (MAB1501R, 1:20,000; Chemicon), RFP (#600401379, 1:2000; Rockland), nuclear factor‐κB (NFκB) (sc‐514451, 1:1000; Santa Cruz), phospho‐NFκB (sc‐136548, 1:1000, Santa Cruz), glyceraldehyde 3‐phosphate dehydrogenase (2118S, 1:2000; Cell Signaling), and TAB1 (67020‐1‐Ig, 1:10.000; Proteintech) and secondary antibodies (goat anti‐mouse [#926‐32210] and goat anti‐rabbit [#926‐68021], all 1:15,000, LI‐COR). Blots were quantified using LI‐COR Odyssey Scanner and Odyssey 3.0 software.

van Woerden G.M., Senden R., de Konink C., Trezza R.A., Baban A., Bassetti J.A., van Bever Y., Bird L.M., van Bon B.W., Brooks A.S., Guan Q., Klee E.W., Marcelis C., Rosado J.M., Schimmenti L.A., Shikany A.R., Terhal P.A., Nicole Weaver K., Wessels M.W., van Wieringen H., Hurst A.C., Gooch C.F., Steindl K., Joset P., Rauch A., Tartaglia M., Niceta M., Elgersma Y, & Demirdas S. (2022). The MAP3K7 gene: Further delineation of clinical characteristics and genotype/phenotype correlations. Human Mutation, 43(10), 1377-1395.

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Antibody-based protein detection protocol

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Antibodies against p-Akt (sc-7985-R), Akt (sc-1618), PI3K (sc-67306), NF-kB (sc-514451) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, U.S.A.). Antibodies specific for LC3 (ab51520), gasdermin D (GSDMD; ab210070), IL18 (ab191152), IL1b (ab9722) were purchased from Abcam (Cambridge, MA, U.S.A.). The actin antibody (ab-2768234) was produced by ABclonal (Wuhan, China). The dilution ratio for all primary antibodies was 1:2000. The secondary antibodies used in the present study were peroxidase AffiniPure goat anti-rabbit-IgG (H+L) (BF03008X) and goat anti-mouse-IgG (H+L) (BF03001X), which were obtained from Bio Dragon (Beijing, China). Secondary antibodies were used at 1:5000 dilution.

Tong W., Guo J, & Yang C. (2020). Tanshinone II A enhances pyroptosis and represses cell proliferation of HeLa cells by regulating miR-145/GSDMD signaling pathway. Bioscience Reports, 40(4), BSR20200259.

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SDS-PAGE and Western Blot Analysis

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Tissue proteins were separated on a 10% sodium dodecyl sulfate polyacrylamide gel and transferred to polyvinylidene difluoride (PVDF) membrane. Membranes were then probed with antibodies from Santa Cruz against nuclear factor-ĸβ (SC-514451), interleukin-6 (SC-57315), C reactive protein (SC-69770), and β-actin ((C4) sc-47778).

Li J., Canseco D., Wang Y., Vale G., Chaudhary J., Anwar A., Baniasadi H., Williams N.S., Gopal P., Sutphin P.D., McDonald J.G., Putnam W.C, & Corbin I.R. (2020). Assessing the Safety of Transarterial Locoregional Delivery of Low-Density Lipoprotein Docosahexaenoic Acid Nanoparticles to the Rat Liver. European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V, 158, 273-283.

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