Mouse aortic samples and MOVAS cell extracts were quantitated by a BCA kit (Thermo Scientific, USA) and subjected to Western blot measurement. The primary antibodies incubation was carried out at 4 °C overnight, followed by secondary antibodies at room temperature for 1 h. The protein expression was detected by Odyssey infrared imaging system (LICOR, USA) and analyzed by Quantity One software. The protein expression was normalized to corresponding GAPDH or Tubulin expression. In our work, the primary antibodies of GSDMD (ab209845), Caspase-11 (ab180673) and α7nAChR (ab10096), were purchased from abcam (UK). The primary antibodies of NLRP3 (#15101), ASC (#67824), Bcl-2 (#2870) and IL-1β (#31202) were purchased from Cell Signaling Technology (USA). Cleaved Capsase 1 (sc-56036) and OPN (Osteopontin, sc-21742) antibodies were obtained from Santa Cruz (USA). α-SMA (14395-1-AP) and IL-18 (10663-1-AP) antibodies were purchased from proteintech (USA). Tubulin (AF0001), Bax (AF0054) and GAPDH (AG019) antibodies from Beyotime (China) were used.
Il 18 10663 1 ap
Manufactured by Proteintech
Sourced in United States, United Kingdom
IL-18 (10663-1-AP) is a primary antibody product offered by Proteintech. This antibody is designed to detect the Interleukin-18 (IL-18) protein. IL-18 is a cytokine that plays a role in immune system regulation.
Automatically generated - may contain errors
Lab products found in correlation
Bca kit, by Thermo Fisher Scientific (3 mentions)
α7nachr ab10096, by Abcam (1 mentions)
α sma 14395 1 ap, by Proteintech (1 mentions)
Gapdh ag019, by Beyotime (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Asc 10500 1 ap, by Proteintech (1 mentions)
Ecl chemiluminescence substrate, by Biosharp (1 mentions)
Il1β 16806 1 ap, by Proteintech (1 mentions)
Nlrp3 19771 1 ap, by Proteintech (1 mentions)
Bca kit, by Biosharp (1 mentions)
Gapdh, by Abcam (1 mentions)
Enhanced chemiluminescence, by Thermo Fisher Scientific (1 mentions)
Cleaved gsdmd, by Abcam (1 mentions)
Rxfp1, by Santa Cruz Biotechnology (1 mentions)
Radioimmunoprecipitation assay buffer, by Beyotime (1 mentions)
3 protocols using il 18 10663 1 ap
1
Comprehensive Protein Analysis in Mouse Aortic Samples
2
Protein Extraction and Western Blot Analysis
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To extract the total protein in cells and tissues, RIPA lysis buffer was employed. Protein was quantified in each group using the BCA kit (Biosharp, Anhui, China) and subsequently mixed with sodium dodecyl sulfate polyacrylamide gel electrophoresis buffer; the protein was adsorbed on the polyvinylidene fluoride membrane. Then, the membranes were blocked in 5% bovine serum albumin (BSA) for 1 hour. ASC (10500-1-AP; Proteintech, Rosemont, IL, USA), cleaved caspase-1 (24232; CST, Danvers, MA, USA), GSDMD-N (ab215203; Abcam, Cambridge, UK), NLRP3 (19771-1-AP; Proteintech), IL-18 (10663-1-AP; Proteintech), IL-1β (16806-1-AP; Proteintech), PTEN (9188; CST), and GAPDH (AB9485; Abcam) primary antibodies were used for overnight incubation. We then incubated horseradish peroxidase (HRP)-labeled secondary antibody IgG-HRP (BL003A; BioSharp). Exposure was performed using an ECL chemiluminescence substrate (Biosharp). To measure protein expressions, GAPDH was used as an internal reference.
3
Western Blot Analysis of Kidney Protein Expression
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Cells or homogenized kidney tissues were harvested in radioimmunoprecipitation assay buffer (Beyotime) and fully lysed on ice. After centrifugation (12 000 rpm, 10 minutes), the supernatants were collected, and the protein concentration was determined. Equal amounts of proteins were separated by SDS‐PAGE and then transferred onto poly(vinylidene fluoride) (PVDF) membranes. The membranes were blocked with 5% nonfat milk/Tris buffered saline with Tween (TBST) and successively incubated with the corresponding primary and secondary antibodies. The protein signals were then assessed using enhanced chemiluminescence (Thermo Fisher Scientific). The following primary antibodies were used: ASC (sc‐514414), PKA (sc‐390548), P2X7R (sc‐514962) and RXFP‐1 (sc‐293228) from Santa Cruz Biotech; cleaved caspase‐1 (4199/89332) and pro‐caspase‐1 (3866) from Cell Signaling Technology; IL‐1β (WL00891) and NLRP3 (WL02635) from Wanleibio (Wanleibio); cleaved GSDMD ([ab215203; Abcam], [sc‐393656; Santa Cruz Biotech]); IL‐18 (10663‐1‐AP; ProteinTech); and glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) (AC002; ABclonal). Peroxidase‐conjugated secondary antibodies (ZSGB‐BIO) were used.
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