Primary macrophages were lysed using radioimmunoprecipitation assay (RIPA) lysis buffer (89900, Pierce, Thermo Fisher Scientific). Protein concentration was quantified using Bradford kit (23236, Pierce). Proteins were then separated by SDS-PAGE electrophoresis and transferred onto a polyvinylidene fluoride (PVDF) membrane. After blocking with 5% non-fat milk, the blots were incubated with primary antibodies at 4°C overnight and a goat anti-mouse HRP-conjugated secondary antibody (31430, Invitrogen; 1:5,000) or goat anti-rabbit HRP-conjugated secondary antibody (31460, Invitrogen; 1:5,000). Signal was visualized using enhanced chemiluminescence (ECL) detection (RPN2105, GE Healthcare). The primary antibodies used in western blot: anti-FABP4 (#2120; 1:1,000), anti-NLRP3 (#15101; 1:1,000), anti-ASC (#67824; 1:1,000), anti-caspase-1 (#3866; 1:1000), anti-cleaved caspase-1 (#4199; 1:1,000), anti-p-p65 (#3033; 1:1,000), anti-p65 (#8242; 1:1000), anti-p-IKKβ (#2697; 1:1,000), anti-IKKβ (#8943; 1:1,000), anti-p-ERK (#4370; 1:2,000), anti-ERK (#4695; 1:1000), anti-p-p38 (#4511; 1:1,000), anti-p38 (#8690; 1:1,000), anti-p-PERK (#3179; 1:1,000), anti-PERK (#5683; 1:1,000), anti-p-eIF2a (#3398; 1:1,000), anti-eIF2a (#5324; 1:1000), and anti-CHOP (#2895; 1:1,000) were obtained from Cell Signaling Technologies (CST, Beverly, MA, USA). Anti-β-actin (ab8227; 1:2,000) was from Abcam.
Anti p ikkβ
Manufactured by Cell Signaling Technology
Anti-p-IKKβ is an antibody used to detect the phosphorylated form of the IKKβ (inhibitor of kappa B kinase beta) protein. IKKβ is a key regulator of the NF-κB signaling pathway, which plays a role in various cellular processes such as inflammation, immune response, and cell survival. This antibody can be used to study the activation and regulation of the NF-κB pathway.
Automatically generated - may contain errors
Lab products found in correlation
Anti ikkβ, by Cell Signaling Technology (2 mentions)
Anti eif2a, by Cell Signaling Technology (1 mentions)
Anti p eif2a, by Cell Signaling Technology (1 mentions)
Anti erk, by Cell Signaling Technology (1 mentions)
Anti p erk, by Cell Signaling Technology (1 mentions)
Anti p p38, by Cell Signaling Technology (1 mentions)
Anti p38, by Cell Signaling Technology (1 mentions)
Anti β actin, by Abcam (1 mentions)
Anti p perk, by Cell Signaling Technology (1 mentions)
Anti chop, by Cell Signaling Technology (1 mentions)
Anti p65, by Cell Signaling Technology (1 mentions)
Anti p84, by Abcam (1 mentions)
Anti p iκbα, by Cell Signaling Technology (1 mentions)
Anti α tubulin, by Santa Cruz Biotechnology (1 mentions)
Bradford assay, by Thermo Fisher Scientific (1 mentions)
Anti iκbα, by Cell Signaling Technology (1 mentions)
2 protocols using anti p ikkβ
1
Western Blot Analysis of Macrophage Signaling
2
Protein Analysis of Cellular Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells and tissues were lysed with RIPA Lysis Buffer, and cleared by centrifugation at 4 °C. The protein concentration of each lysate was determined with the Bradford assay (Thermo Scientific, Massachusetts, USA). Equal amounts of protein were separated by SDS-PAGE gels, and then transferred to polyvinylidene difluoride membrane, and probed with corresponding antibodies: Fezf2 (Abcam, cat. no. ab69436), anti-p-IκBα (Cell Signaling Technology, cat. no. 2859), anti-IκBα (Cell Signaling Technology, cat. no. 4814), anti-p-IKKβ (Cell Signaling Technology, cat. no. 2078), anti-IKKβ (Cell Signaling Technology, cat. no. 2370), anti-p65 (Cell Signaling Technology, cat. no. 4764), anti-p84 (Abcam, cat. no. ab131268), and anti-α-tubulin (Santa Cruz, cat. no. sc-5286).
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