Caspase 1

Manufactured by GeneTex

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Caspase 1 is a protein-cleaving enzyme that plays a key role in the inflammatory response and programmed cell death. It is involved in the activation of pro-inflammatory cytokines and the initiation of the apoptotic pathway.

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4 protocols using caspase 1

Protein Expression Analysis by Western Blot

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Western blots performed with 10, 12, or 15% SDS-polyacrylamide gels were incubated with primary antibodies for IL-1ß (R&D Systems, Minneapolis, MN), NLRP3 (LSBio, Seattle, WA), Ogg1 (Novus Biologicals, Littleton, CO), Caspase 1 (Genetex, Irvine, CA), and DNMT3b (Abcam, Cambridge, MA). Western blots were visualized using alkaline phosphatase-conjugated secondary antibodies (Sigma-Aldrich).

Haldar S., Dru C., Mishra R., Tripathi M., Duong F., Angara B., Fernandez A., Arditi M, & Bhowmick N.A. (2016). Histone deacetylase inhibitors mediate DNA damage repair in ameliorating hemorrhagic cystitis. Scientific Reports, 6, 39257.

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Renal Protein Extraction and Immunoblotting

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Cytoplasmic protein was extracted from renal tissues and cultured cells and immunoblotting using antibodies against mouse NLRP3, caspase‐1 (GeneTex), IL‐1β (R&D Systems, Minneapolis, MN, USA), IL‐18 (Santa cruz), NADPH p47^phox (Santa Cruz), NQO1 (Abcam, Bristol, UK), ATG5 (MBL, Japan), LC3B (GeneTex), p62 (Santa cruz), SIRT1 (Cell Signaling), SIRT3 (Cell Signaling), p‐ERK (Cell Signaling), p‐JNK (Cell Signaling), p‐p38 (Cell Signaling) and horseradish peroxidase (HRP)‐conjugated IgG antibodies (Santa cruz) as described previously.⁹, ¹⁰ β‐actin (Santa Cruz) was used as an internal control for cytosolic protein.

Wu C., Hua K., Yang S., Tsai Y., Yang S., Hsieh C., Wu C., Chang J., Arbiser J.L., Chang C., Chen A, & Ka S. (2020). Tris DBA ameliorates IgA nephropathy by blunting the activating signal of NLRP3 inflammasome through SIRT1‐ and SIRT3‐mediated autophagy induction. Journal of Cellular and Molecular Medicine, 24(23), 13609-13622.

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Alpinumisoflavone Inhibits Inflammation via TLR4/NLRP3 Pathway

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Alpinumisoflavone (AIF) was purchased from BJYM Pharmaceutical & Chemical Co., Ltd. (Beijing, People's Republic of China). The purity of AIF used in the present study was >95%. LPS (Escherichia coli 055:B5), dexamethasone (Dex), thiazolyl blue tetrazolium bromide (MTT), 2′,7′-dihydrofluorescein diacetate (DCFH-DA), dimethyl sulfoxide (DMSO) and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against TLR-4, NLRP3, Hsp90α, TRAF6, COX-2, Act1, CAT, phospho-JNK, GPx, SOD, caspase-1, and IL-1β were purchased from Gene Tex (San Antonio, TX, USA). Antibodies against iNOS, NF-κB, IκBα, p38, HO-1, ICAM-1 and β-actin were purchased from Abcam (Cambridge, UK, USA). Antibodies against Phospho-IKKα/β, IKKβ, JNK, Phospho-ERK, ERK, Phospho-p38, and Phospho-IκB-α were purchased from Cell Signaling Technology (Beverly, MA, USA). Polyvinylidene fluoride transfer membranes (Immobilon P) and enhanced chemiluminescence (ECL) reagent were from EMD Millipore (Bedford, MA, USA). The protein assay dye reagent were purchased from Bio-Rad (Hercules, CA, USA).

Li P.Y., Liang Y.C., Sheu M.J., Huang S.S., Chao C.Y., Kuo Y.H, & Huang G.J. (2018). Alpinumisoflavone attenuates lipopolysaccharide-induced acute lung injury by regulating the effects of anti-oxidation and anti-inflammation both in vitro and in vivo. RSC Advances, 8(55), 31515-31528.

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Western Blot Analysis of Cardiac Markers

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Total protein samples were extracted from tissues or cells using the same procedure previously described [36 (link)]. Briefly, heart tissues or cardiomyocytes were lysed by RIPA buffer. After centrifugation, the supernatant was collected. Protein extracts were mixed with SDS‐PAGE. After electrophoresis, the protein transferred onto a nitrocellulose membrane. Then, the membranes were blocked and incubated with the primary antibodies: BNP (1:1000, ABclonal, China), β-MHC (1:1000, ABclonal, China), Caspase-1 (1:1000, GeneTex, USA), Nrf2 (1:1000, Proteintech, China)，HO-1 (1:1000, Wanleibio, China)，NLRP3 (1:1000, ABclonal, China)，Gasdermin-D (ABclonal, China), USP14 (1:1000, Proteintech, China), Snail (1:1000, Proteintech, China)，Twist (1:1000, Proteintech, China)，Slug (1:1000, ABclonal, China), CD31 (1:1000, ABclonal, China), VE-cadherin (1:1000, ABclonal, China), α-SMA (1:1000, BOSTER, China), Keap1 (1:1000, Proteintech, China), type I collagen (1:1000, ABclonal, China), type Ⅲ collagen (1:1000, affinity, USA).

Ba L., E M., Wang R., Wu N., Wang R., Liu R., Feng X., Qi H., Sun H, & Qiao G. (2024). Triptolide attenuates cardiac remodeling by inhibiting pyroptosis and EndMT via modulating USP14/Keap1/Nrf2 pathway. Heliyon, 10(2), e24010.

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