The protein of hippocampus tissue was prepared using tissue isolation kit and measured using BCA kit (Beyotime Biotechnology, Jiangsu, China). Samples of protein (15 mg) were subjected to dodecyl sulfate, sodium salt-polyacrylamide gel electrophoresis and were transferred onto polyvinylidene fluoride membranes. Membranes were blocked and incubated with a set of primary antibodies (anti-p-IKKα, anti-IKKα/β, anti-IκBα, and anti-p-IκBα, Bioss Inc., USA) overnight at 4°C. The blots were visualized using an ECL detection kit (GE Healthcare, Germany). A densitometric analysis was performed.
Anti p iκbα
Manufactured by Bioss Antibodies
Sourced in China
Anti-p-IκBα is a laboratory reagent that detects the phosphorylated form of the inhibitor of NF-κB alpha (IκBα) protein. IκBα is a key regulator of the NF-κB signaling pathway, and its phosphorylation is a critical step in the activation of NF-κB. This antibody can be used to study the activation of the NF-κB pathway in various cellular and experimental contexts.
Automatically generated - may contain errors
Lab products found in correlation
Anti iκbα, by Bioss Antibodies (2 mentions)
Ecl detection kit, by GE Healthcare (1 mentions)
Bca kit, by Beyotime (1 mentions)
Anti p65, by Proteintech (1 mentions)
Histone h3, by Abcepta (1 mentions)
Anti e cadherin, by Proteintech (1 mentions)
Anti n cadherin, by Proteintech (1 mentions)
Ecl reagent, by Beyotime (1 mentions)
Anti β actin, by Keygen Biotech (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions)
Nuclear and cytoplasmic protein extraction kit, by Beyotime (1 mentions)
Anti fibronectin, by Proteintech (1 mentions)
F4 80, by Abcam (1 mentions)
Anti p p65, by Santa Cruz Biotechnology (1 mentions)
Bs 1287r, by Bioss Antibodies (1 mentions)
Bs 2513r, by Bioss Antibodies (1 mentions)
Alanine transaminase alt, by Nanjing Jiancheng (1 mentions)
Aspartate aminotransferase ast kit, by Nanjing Jiancheng (1 mentions)
4 protocols using anti p iκbα
1
Hippocampus Protein Analysis by Western Blot
2
Protein Expression Analysis in CRC and Liver Metastases
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CRC cells or liver metastatic tissues were lysed with RIPA Lysis Buffer (Beyotime, China) containing 1% PMSF (Beyotime). Besides, the nuclear protein was isolated by the Nuclear and Cytoplasmic Protein Extraction Kit (Beyotime). Protein from different conditions were separated by 6–14% SDS-PAGE and then transferred to PVDF membranes. After blocking with 5% non-fat milk or 1% BSA, the membranes were incubated with following primary antibodies at 4 °C overnight: anti-E-cadherin (1:1000, 60,335–1-Ig, Proteintech, China), anti-N-cadherin (1:1000, 66,219–1-Ig, Proteintech, China), anti-fibronectin (1:500, 15,613–1-AP, Proteintech, China), anti-p-IκBα (1:500, bs-2513R, Bioss, China), anti-IκBα (1:500, bs-1287R, Bioss, China), anti-p65 (1:500, 10,745–1-AP, Proteintech, China), anti-β-actin (1:500, KGAA001, KeyGen, China) and Histone H3 (1:2000, AM8433, ABGENT, USA). After rinsed with TSBT, the membranes were incubated with their corresponding secondary antibodies at 37 °C for 45 min. The protein bands were visualized using the ECL reagent (Beyotime).
3
Immunohistochemical Analysis of Liver Inflammation
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Livers were fixed with 10% buffered formalin and embedded in paraffin according to the standard protocol. Paraffin-embedded liver tissue was cut into 5-μm-thick sections. After dewaxing and hydration, antigen retrieval was conducted with 0.01 M sodium citrate buffer solution (pH 6.0). Sections were incubated with anti-p-IκBα (1:200 dilution, Bioss, catalog [cat.] no. 5515R), anti-p-p50 (1:200 dilution, Bioworld, cat. no. 4132), anti-p-p65 (1:200 dilution, Santa Cruz, cat. no. 136548), and F4/80 (1:100 dilution, Abcam, cat. no. 111101) monoclonal antibodies at 4°C overnight. The color reaction was developed with an HRP-linked polymer detection system and counterstaining with hematoxylin was performed before dehydrating in graded alcohols. F4/80, TGR5-, p-IκBα-, p-p50-, and p-p65-positive cells were counted in ten randomly selected fields from each slide, at a magnification of ×200.
4
Quantitative Immunoassay Protocol
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Alanine transaminase (ALT) and aspartate aminotransferase (AST) kits were purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd. (Nanjing, China). Immunoglobulin G (IgG) and secretory immunoglobulin A (sIgA) detection kits were purchased from Shanghai Enzyme Linked Biotechnology Co., Ltd.(Shanghai, China). The primary antibodies used in this study were as follows: anti−p−IκBα (1:1000, bs−2513R, Bioss (Beijng, China)), anti−IκBα (1:1000, bs−1287R, Bioss), β−actin (1:2000, 20536−1−AP, Proteintech (Wuhan, China)), anti−p65 (1:1000, 10745−1−AP, Proteintech), anti−TRAF6 (1:5000, 66498−1−Ig, Proteintech), anti−IRAK1 (1:1000, 10478−2−AP, Proteintech), and anti−p−p65 (1:500, AP3749a, Abcepta (Suzhou, China)). Among the primary antibodies used in Western blotting, only anti-TRAF6 (clone number: 1D1E1) was a murine monoclonal antibody, while the remaining were rabbit polyclonal antibodies.
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