Sc 7178

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Sc-7178 is a lab equipment product offered by Santa Cruz Biotechnology. It serves as a core tool for researchers in life science and biotechnology fields. The detailed specifications and intended use of this product are not available at this time.

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Lab products found in correlation

Sc 2040, by Santa Cruz Biotechnology (1 mentions) Goat anti rabbit igg biotin conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions) Sc 1616, by Santa Cruz Biotechnology (1 mentions) Sc 33020, by Santa Cruz Biotechnology (1 mentions) Sc 372, by Santa Cruz Biotechnology (1 mentions) Iκbα sc 371, by Santa Cruz Biotechnology (1 mentions) Ikk inhibitor bay 117085, by Cayman Chemical (1 mentions) Bortezomib, by Selleck Chemicals (1 mentions) Ab46540, by Abcam (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) Sc 7151, by Santa Cruz Biotechnology (1 mentions) Odyssey infrared emsa kit, by LI COR (1 mentions) Wbkls0500, by Merck Group (1 mentions) Ab16045, by Abcam (1 mentions) Ab45036, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Rabbit anti-p50 (sc-7178, (2 citations) Rabbit anti-NF-κB p50 (sc-7178, (2 citations)

5 protocols using sc 7178

Immunohistochemical Analysis of Hepatic Tissues

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Hepatic tissues were fixed in 10% buffered neutral formalin, washed, dehydrated, cleared, embedded in paraffin, cast then sectioned. Tissue sections were deparaffinized and treated with 3% H₂O₂ for 10 min to inactivate the peroxidases. Subsequently, samples were heated in 10 mM citrate buffer at 121°C for 30 min for antigen retrieval and blocked in 5% normal serum for 20 min, and pancreas was incubated with a rabbit polyclonal anti-glutathione primary antibody (1:100; sc-71155; Santa Cruz Biotechnology, Inc., Dallas, TX) or NFκB p50 antibody (1:100; sc-7178; Santa Cruz Biotechnology, Inc.) in PBS overnight at 4°C. After three extensive washes with PBS, the sections were incubated with a goat anti-rabbit IgG biotin-conjugated secondary antibody (1:2,000; sc 2040; Santa Cruz Biotechnology, Inc.) for 20 min at 32°C. After further incubation with horseradish peroxidase-labeled streptavidin, antibody binding was visualized using diaminobenzidine, and the sections were counterstained with hematoxylin.

Alkhedaide A.Q., Ismail T.A., Alotaibi S.H., Nassan M.A, & Shehri Z.S. (2018). Preventive effect of artemisinin extract against cholestasis induced via lithocholic acid exposure. Bioscience Reports, 38(6), BSR20181011.

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Protein Expression and Purification Protocols

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pRC-HA-hRelA (1–551), His-p50 (1–435), GST-RelA-AD (325–551), and GST-RelA-AD (429–551) have been previously described.^{23 (link)} Wild-type RelA was subcloned into a plasmid to express RelA as a Flag fusion protein. The HA-RelA construct was used as the template to generate a RelA-RHR fragment and insert it into the pET15b plasmid that expresses protein as N-terminal His fusion protein. All Flag-p53 constructs were a kind gift from S. Lauberth and were used as the template to amplify the inserts and subclone them into the pET15b His-tagged plasmid. The p53-AD (1–95) and p53-DBD (96–292) plasmids were generated using the His-p53 wild-type (wt) construct as the template followed by their subcloning into the pET24d His-tagged plasmid. His-RPS3, His-OGG1, and His-HMGA1 [formerly HMGI(Y)] plasmids were a kind gift obtained from M. Lenardo and T. Hazra. E-Selectin and HIV luciferase reporters containing the specific κB DNA promoter have been previously described.^{23 (link)} The antibodies recognizing RelA (sc-372), p50 (sc-7178), and actin (sc-1616) were purchased from Santa Cruz. Anti-HA.11 (901501) was obtained from BioLegend. Anti-p84 (C1C3) was purchased from GeneTex. Anti-His, anti-p53, and anti-tubulin antibodies and TNF-α were gifts from BioBharati Life Science (Kolkata, India).

Mulero M.C., Shahabi S., Ko M.S., Schiffer J.M., Huang D.B., Wang V.Y., Amaro R.E., Huxford T, & Ghosh G. (2018). Protein Cofactors Are Essential for High-Affinity DNA Binding by the Nuclear Factor κB RelA Subunit. Biochemistry, 57(20), 2943-2957.

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Characterization of NF-κB signaling components

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Purified polyclonal antibodies against human p65 NFκB (sc-372), phosphorylated p65 NFκB at S536 (sc-33020), p50 NFκB (sc-7178), and IκBα (sc-371) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Purified polyclonal antibody against actin was from Sigma (St. Louis, MO, USA). Horseradish peroxidase (HRP)-conjugated anti-rabbit, anti-mouse and anti-goat secondary antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA). Bortezomib was obtained from Selleck Chemicals (Houston, TX, USA). The IKK inhibitor Bay 117085 was purchased from Cayman Chemicals (Ann Arbor, MI, USA). All other reagents were molecular biology grade and were from Sigma (St. Louis, MO, USA).

Singha B., Gatla H.R., Phyo S., Patel A., Chen Z.S, & Vancurova I. (2015). IKK inhibition increases bortezomib effectiveness in ovarian cancer. Oncotarget, 6(28), 26347-26358.

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EMSA Analysis of NF-κB Subunits

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Nuclear extracts for EMSA were prepared as described previously from HEK 293T cells transfected with pcDNA3.1-EGFP-p50 and pcDNA3.1-EGFP-p65. EMSA 5′ IRDye700 oligonucleotides were obtained from Integrated DNA Technologies (IDT) and annealed to obtain double-stranded DNA probes. For the oligo and probe sequences used in the EMSAs, see Figure 2C, lower panel. The binding reactions were performed as described, using the Odyssey Infrared EMSA kit (LI-COR Biosciences, Lincoln, NE, USA). Oligonucleotides in 1300-fold molar excess were used for competition experiments. The super-shift experiments were performed with 2 μg of rabbit anti-p50 (sc-7178, Santa Cruz), rabbit anti-p65 (sc-7151, Santa Cruz), or rabbit control IgG (ab46540, Abcam). The reaction mixtures were run through 5% native polyacrylamide gels for 100 min in 0.5× Tris/Borate/EDTA (TBE) buffer. The gels were scanned immediately with an Odyssey^® Infrared Imaging System (LI-COR Biosciences).

Verhoeven R.J., Tong S., Zong J., Chen Y., Tsao S.W., Pan J, & Chen H. (2018). NF-κB Signaling Regulates Epstein–Barr Virus BamHI-Q-Driven EBNA1 Expression. Cancers, 10(4), 119.

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Whole Cell Protein Extraction and Western Blot Analysis

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For whole cell extractions, liver samples were homogenized in lysis buffer (50mM Tris-HCl pH Remaining cell debris were removed by centrifugation (24,000 g, 20 min). Protein concentration was assessed by Bradford assay [15] . Proteins (20 µg) were separated on SDS-PAGE and transferred on PVDF membrane. Blocking was performed using 5% milk, 1h at room temperature. Membranes were then incubated with primary antibodies overnight at 4 o C.
Dilutions of the primary antibodies were of 1:50,000 for cyclophilin B (Abcam, ab16045), 1:1000 for PPARγ (Abcam, ab45036), SREBP-1 (Santa Cruz, sc13551) FASN (Abcam, ab22759) and NF-κB p50 (Santa Cruz, sc7178). Primary antibodies were detected using goat anti-rabbit HRP conjugated IgG antibodies (Cell Signaling Tech., 7074S) at 1:1000 and visualized by using chemiluminescent HRP substrate (Millipore, WBKLS0500). Amidoblack staining was used as a loading control. Briefly, membranes were stained for 20 min in amidoblack solution (0.1% amidoblack, 40% v/v methanol and 10% v/v acetic acid) and washed 10 min twice in decolouration solution (40% v/v methanol and 10% v/v acetic acid). Bands were quantified by densitometry using Image J software.

Desmarais F., Bergeron K.F., Rassart E, & Mounier C. (2019). Apolipoprotein D overexpression alters hepatic prostaglandin and omega fatty acid metabolism during the development of a non-inflammatory hepatic steatosis. Biochimica et biophysica acta. Molecular and cell biology of lipids, 1864(4).

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