Anti-NLRP3 (15101), anti-ASC (67824), and anti-Ub (3936) were purchased from Cell Signaling Technology (MA, USA). Anti-IL-1β (ab9722) was purchased from Abcam (Cambridge, UK). Anti-Myc (AE070), anti-HA (AE008), anti-Flag (AE004), anti-Caspase-1 (A0964) and anti-YOD1 (A13270) were purchased from ABclonal (Wuhan, China). Anti-β-actin (T0022) was purchased from Affinity Biosciences (State of Ohio, USA). Protein A/G agarose used for immunoprecipitation was purchased from Thermo Fisher Scientific (Waltham, USA). Mouse D-Dimer ELISA Kit (E-EL-M0400c), Mouse thrombin-antithrombin (TAT) ELISA Kit (E-EL-M1138c), Mouse interleukin 1 Beta (IL-1β) ELISA Kit (E-EL-M0037c), Mouse plasminogen activator inhibitor 1 (PAI-1) ELISA Kit (E-EL-M3041) were purchased from Elabscience (Wuhan, China). Fibrinogen (FIB) ELISA Kit (JL20600) was purchased from Jianglai Biotechnology (Shanghai, China). MCC950 Sodium (S7809) was purchased from Selleck (TX, USA).
Anti il 1β ab9722
Manufactured by Abcam
Sourced in United Kingdom
Anti-IL-1β (ab9722) is a primary antibody that recognizes the interleukin-1 beta (IL-1β) protein. IL-1β is a pro-inflammatory cytokine involved in various immune and inflammatory responses. This antibody can be used to detect and quantify IL-1β in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Protein a g agarose, by Thermo Fisher Scientific (1 mentions)
Mouse d dimer elisa kit, by Elabscience (1 mentions)
Mouse interleukin 1 beta il 1β elisa kit, by Elabscience (1 mentions)
Anti ub 3936, by Cell Signaling Technology (1 mentions)
Ripa buffer reagent, by Thermo Fisher Scientific (1 mentions)
Anti atg5 a0856, by Merck Group (1 mentions)
Anti p62 p0067, by Merck Group (1 mentions)
Anti gapdh af7021, by Affinity Biosciences (1 mentions)
Anti nlrp3, by Cell Signaling Technology (1 mentions)
Anti caspase3 sc 56053, by Santa Cruz Biotechnology (1 mentions)
Anti caspase 1, by Abcam (1 mentions)
Gapdh, by GeneTex (1 mentions)
Anti ho 1, by Bioss Antibodies (1 mentions)
4 protocols using anti il 1β ab9722
1
Molecular Mechanisms of NLRP3 Inflammasome Activation
2
Autophagy Markers Quantification in Corneal Cells
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To determine the change in autophagy, we detected the expression of the autophagy-associated markers LC3II/I and P62 by Western blotting. Protein samples from mouse corneal stromal cells or corneal tissues were extracted using RIPA buffer reagent (Thermo Fisher Scientific, Germany) and separated using 12.5% SDS-PAGE gels. After electrophoresis, the separated proteins were transferred to a polyvinylidene difluoride (PVDF) membrane and blocked with 5% defatted milk powder (Applygen, Beijing, China). Primary antibodies, including anti-LC3 I/II (L7543; Sigma, St. Louis, MO, USA), anti-p62 (P0067; Sigma), anti-IL-1β (ab9722; Abcam, Cambridge, UK), anti-ATG5 (A0856; Sigma), and anti-GAPDH (AF7021; Affinity Biosciences, Cincinnati, OH, USA), were incubated overnight at 4°C. After washing with PBS containing 0.05% Tween 20 (Bio-Rad, Hercules, CA, USA) three times, the membranes were incubated with specific secondary antibodies for 2 hours. Finally, the proteins were quantified using the ECL detection reagent (WBKLS0100; Millipore, Billerica, MA, USA).
3
Western Blot Analysis of Liver Proteins
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The Western blot analysis was performed as described previously [33 (link)]. The liver homogenates of each group with equal amounts of protein were separated by SDS-PAGE and transferred to polyvinylidene difluoride membranes. After blocking of the nonspecific binding sites with 5% non-fat dry milk in 15 mM Tris/150 mM NaCl buffer (pH 7.4), the membranes were hybridized with antibodies against anti-NLRP3 (#15101, Cell Signaling Technology, Danvers, MA, USA), anti-caspase-1 (#3019, BioVision Inc, Milpitas, CA, USA), anti-IL-1β (ab9722, Abcam, Cambridge, UK), anti-caspase 3 (SC-56053, Santa Cruz Biotechnology, CA, USA), anti-pi class GSH-S-transferase (PGST) (610719, BD Biosciences, San Jose, CA, USA), anti-heme oxygenase-1 (HO-1) (374090, Merck Millipore, Billerica, MA, USA), and GAPDH (GTX100118 GeneTex Inc, Irvine, CA, USA).
4
Western Blot Analysis of Cellular Signaling
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After cell lysates were obtained from control and agent-treated cells, approximately 40 μg proteins were subjected to 10% SDS–page and transferred to nitrocellulose membrane. Membranes were blocked at room temperature for 1 h in blocking buffer containing 5% nonfat dry milk to prevent nonspecific binding and then incubated with primary antibodies overnight at 4°C. The primary antibodies used in the present study was anti-Nrf2 (Biorbyt, Cambridge, 1 : 200), anti-NLRP3 (MyBioSource, LLC, San Diego, California, 1 : 1000, anti-IL-1β (ab-9722, Abcam, 1 : 1000), anti-HO-1 (Bioss Antibodies, Beijing, 1 : 1,000), and Anti-NQO1 (Bioss Antibodies, 1 : 1,000). The membranes were washed in TBST (50 mmol/L Tris-HCl, pH 7.6; 150 mmol/L NaCl; 0.1% Tween 20) for 30 min and incubated with an appropriate secondary antibody (Sigma, 1 : 2000 dilution). Bound antibody was visualized with a commercial enhanced chemiluminescence kit (Amersham Pharmacia Biotech) and Kodak film. The level of each protein was expressed relative to the amount of actin.
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