Ikkα β sc 7607

Manufactured by Santa Cruz Biotechnology

Sourced in United States

IKKα/β (sc-7607) is a mouse monoclonal antibody that recognizes the IKKα and IKKβ subunits of the IKB kinase (IKK) complex. The IKK complex is a key regulator of the NF-κB signaling pathway.

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Lab products found in correlation

Iblot dry blotting system, by Thermo Fisher Scientific (1 mentions) Phospho ikbα, by Cell Signaling Technology (1 mentions) Ez ecl chemiluminescence detection kit, by Sartorius (1 mentions) Erk1 2, by Cell Signaling Technology (1 mentions) P erk1 2, by Cell Signaling Technology (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) P iκbα, by Cell Signaling Technology (1 mentions) Gapdh, by Bioworld Technology (1 mentions) Interleukin 6 (il 6), by Nanjing Jiancheng (1 mentions) Ab22048, by Abcam (1 mentions) Hydroxyproline testing kit a030 2, by Nanjing Jiancheng (1 mentions) Tgf β1, by Nanjing Jiancheng (1 mentions) Ab5694, by Abcam (1 mentions) Tnf α, by Nanjing Jiancheng (1 mentions) Lipopolysaccharides (lps), by Nanjing Jiancheng (1 mentions) Mcp 1, by Nanjing Jiancheng (1 mentions) Enhanced chemiluminescence reagent, by Bio-Rad (1 mentions) Glyceraldehyde 3 phosphate dehydrogenase, by Santa Cruz Biotechnology (1 mentions) Phospho jnk 4668, by Cell Signaling Technology (1 mentions) Iκb sc 371, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

IKKα/β (16 citations) Sc-7607 (4 citations)

6 protocols using ikkα β sc 7607

Quantification of IKK and IκB Proteins

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Total protein was separated by SDS-PAGE and transferred onto PVDF membranes by the iBlot Dry Blotting System (Invitrogen, MA, USA). Blots were incubated overnight at 4 °C with primary anti-human antibodies for phospho-IKK α/β (2697 S, 1:1000; Cell Signaling Technology, MA, USA), IKK α/β (sc-7607,1:400; Santa Cruz Biotechnology, TX, USA) phospho-IKB-α (9246 S, 1:200; Cell Signaling Technology) and IKB-α (sc-203, 1:200; Santa Cruz Biotechnology). Thereafter, the blots were incubated with either donkey anti-rabbit (Biolegend, CA, USA) or anti-mouse (Cell Signaling Technology) HRP-linked antibody (1:2000). To assess housekeeping protein expression, membranes were reblotted overnight at 4 °C with primary rabbit anti-human β-actin HRP conjugate (Cell Signaling Technology). Bands were visualized using the EZ-ECL chemiluminescence detection kit (Biological Industries, Israel). Total IKK α/β, IKB-α and β-actin were used as internal controls when applicable to verify basal level expression and equal protein loading. Further details included as Supplementary Information.

Romo M., López-Vicario C., Pérez-Romero N., Casulleras M., Martínez-Puchol A.I., Sánchez B., Flores-Costa R., Alcaraz-Quiles J., Duran-Güell M., Ibarzábal A., Espert J.J., Clària J, & Titos E. (2022). Small fragments of hyaluronan are increased in individuals with obesity and contribute to low-grade inflammation through TLR-mediated activation of innate immune cells. International Journal of Obesity (2005), 46(11), 1960-1969.

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Diethylnitrosamine-Induced Liver Fibrosis Model

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BBD was obtained from Shanghai Pharmaceuticals Holding Co., Ltd, Shanghai, China, and dissolved in normal saline (NS). Diethylnitrosamine (DEN) was diluted in normal water at a concentration of 1:10,000 (v/v). Hydroxyproline Testing Kit (A030-2) was bought form Jiancheng, Nanjing, China. The antibody of α-smooth muscle actin (α-SMA) (ab5694, Abcam), Toll-like receptor 4 (TLR4) (ab22048, Abcam), pIKKα/β(2697, Cell Signaling), IKKα/β(sc-7607, Santa Cruz), pIκBα(2859, Cell Signaling), IκBα(4812, Cell Signaling), p ERK1/2 (4370, Cell Signaling), ERK1/2 (4695, Cell Signaling), Cyclin D1 (ab134175,abcam) and GAPDH (AP0063, Bioworld) were used for western blot or cellular immunofluorescence staining. LPS (H178, Jiancheng, Nanjing, China), IL-6 (H007, Jiancheng, Nanjing, China), TGF-β1(H034, Jiancheng, Nanjing, China), TNF-α (H052, Jiancheng, Nanjing, China), MCP-1 (H115, Jiancheng, Nanjing, China) were used for endotoxin enzyme-linked immunosorbent assay (ELISA) test. LPS was purchased from Sigma.

Liang L., Yang X., Yu Y., Li X., Wu Y., Shi R., Jiang J., Gao L., Ye F., Zhao Q., Li R., Wei L, & Han Z. (2016). Babao Dan attenuates hepatic fibrosis by inhibiting hepatic stellate cells activation and proliferation via TLR4 signaling pathway. Oncotarget, 7(50), 82554-82566.

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Western Blot Analysis of Renal Signaling Pathways

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Lysate homogenates of renal tissues or cells were prepared, and Western blotting was performed, as previously described unless otherwise indicated.^{49 (link)} Each nitrocellulose membrane was incubated with antibodies to phospho–JNK (4668, 1:1000, Cell Signaling Technology, Danvers, MA), NF-κB p65 (sc-7151, 1:500, Santa Cruz Biotechnology), phospho-IκB (sc-8404, 1:500, Santa Cruz Biotechnology), IκB (sc-371, 1:500, Santa Cruz Biotechnology), IKKα/β (sc-7607, 1:500, Santa Cruz Biotechnology), phospho-IKKα/β (sc-2697, 1:500, Santa Cruz Biotechnology), laminin B (sc-6216, 1:500, Santa Cruz Biotechnology), FGF1 (Abcam, 1:2000), or glyceraldehyde 3-phosphate dehydrogenase (1:5,000, Santa Cruz Biotechnology). The immunoreactive bands were then detected by incubating with secondary antibody (sc-2004, Santa Cruz Biotechnology) conjugated with horseradish peroxidase and visualized using enhanced chemiluminescence reagents (Bio-Rad, Hercules, CA). All Western blots were repeated at least 3 times. The density of the immunoreactive bands was quantified using ImageJ software (NIH, Bethesda, MD). For densitometric quantification of JNK, the total amount of both 46 kD JNK1 and 54kD JNK2 was calculated.

Liang G., Song L., Chen Z., Qian Y., Xie J., Zhao L., Lin Q., Zhu G., Tan Y., Li X., Mohammadi M, & Huang Z. (2017). Fibroblast growth factor 1 ameliorates diabetic nephropathy by an anti-inflammatory mechanism. Kidney international, 93(1), 95-109.

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IGF2 Imprinting and Regulation

Cited 1 time

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RNA was isolated from the cells or mouse prostate tissues using Rneasy Kit (Qiagen) with the addition of Dnase I to minimize DNA contamination. Imprinting was performed using a FluPE assay as previously described [6] (link). For human IGF2, a single nucleotide polymorphism identified on IGF2 exon 7 (G/C) was used to identify individual alleles. IGF2 imprinting was examined on Exon 6 (A/G) in mouse prostate tissues. Differences were determined by calculating the ratio of their respective spectral intensities [repressed allele (Ai)/active allele (Aa)]. Quantitative PCR was performed using an iCycler (Bio-Rad) and SYBR Green PCR master mix (Applied Biosystems) to measure gene expression, primers are available on request. Western blot was performed to detect protein expression using antibodies for CTCF (Cat #07-729, Millipore), NF-κB p50 (4D1, Santa Cruz), NF-κB p65 (c-20, Santa Cruz), NF-κB p100/52 (18D10, cell signaling), IKKα/β (sc-7607, Santa Cruz) and IκBα (c-21, Santa Cruz) and α-Tubulin (DM1A, EMD).

Yang B., Wagner J., Damaschke N., Yao T., Wuerzberger-Davis S.M., Lee M.H., Svaren J., Miyamoto S, & Jarrard D.F. (2014). A Novel Pathway Links Oxidative Stress to Loss of Insulin Growth Factor-2 (IGF2) Imprinting through NF-κB Activation. PLoS ONE, 9(2), e88052.

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Western Blot Analysis of Inflammatory Markers

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Inducible nitric oxide synthase (iNOS; M00368) was used as a primary antibody from Boster Biological Technology (Pleasanton, CA, USA). Inhibitors of κB-α (IκB-α; sc-203), IκB kinase-α/β (IKK-α/β; sc-7607), HO-1 (sc-136960), and β-actin (sc-47778) were used as primary antibodies (Santa Cruz Biotechnology, Inc., Dallas, Texas, USA). NF-κB p65 (4764), phospho-IκB-α (9246), phospho-IKK-α/β (Ser176/180; 2697), kelch-like ECH-associated protein 1 (keap1; sc-514914), and poly ADP-ribose polymerase (PARP; 9542) were used as primary antibodies (Cell Signaling Technology, Danvers, MA, USA). Nrf2 (ab92946) was used as a primary antibody (Abcam, Cambridge, UK). Horseradish peroxidase-conjugated (HRP) secondary antibodies were obtained from Jackson ImmunoResearch (West Grove, PA, USA). Commercial enzyme-linked immunosorbent assay (ELISA) kits for PGE2, myeloperoxidase (MPO), and SOD were purchased from R&D Systems (Minneapolis, MN, USA), Biovision (Milpitas, CA, USA), and Abcam (Cambridge, MA, USA). Unless otherwise specified, all remaining reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Kim H.J., Jin B.R., Lee C.D., Kim D., Lee A.Y., Lee S, & An H.J. (2024). Anti-Inflammatory Effect of Chestnut Honey and Cabbage Mixtures Alleviates Gastric Mucosal Damage. Nutrients, 16(3), 389.

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Comprehensive Western Blotting Assay

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Western blotting was performed as previously described [25 (link)]. Primary antibodies were as follows: pP65 (Ser 536) (sc3020), pP65 (Ser 276) (sc101749), P65 (sc372), COX-2 (cyclooxygenase 2) (sc1747-R), inducible nitric oxide synthase (iNOS) (sc7271), AKT (sc1618), pAKT (Ser 473) (sc7985), inhibitor of kappa B alpha (IKBα) (sc847), pIKBα (Ser32) (sc8404), inhibitor of kappa B kinase alpha/beta (IKKα/β) (sc7607), β-Actin (sc47778), and transcription factor IIB (TFIIB) (sc225) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). pIKKα/β (Ser 176/180) (#2697s) and PTEN and phosphoinositide dependent kinase 1 (PDK1) Ab sample kit (#9652) were purchased from Cell Signaling (Beverly, MA, USA).

Moon K.M., Lee B., Jeong J.W., Kim D.H., Park Y.J., Kim H.R., Park J.Y., Kim M.J., An H.J., Lee E.K., Ha Y.M., Im E., Chun P., Ma J.Y., Cho W.K., Moon H.R, & Chung H.Y. (2017). Thio-barbiturate-derived compounds are novel antioxidants to prevent LPS-induced inflammation in the liver. Oncotarget, 8(53), 91662-91673.

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