Caspase 1

Manufactured by Abcam

Sourced in United States, United Kingdom, China

Caspase-1 is a laboratory tool that plays a central role in the inflammatory response and programmed cell death (apoptosis). It is an enzyme responsible for cleaving and activating other proteins involved in these biological processes.

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Lab products found in correlation

Nlrp3, by Abcam (44 mentions) Il 1β, by Abcam (37 mentions) Il 18, by Abcam (19 mentions) Pvdf membrane, by Merck Group (18 mentions) Gsdmd, by Abcam (17 mentions) Gapdh, by Abcam (15 mentions) Il 1β, by Cell Signaling Technology (12 mentions) β actin, by Abcam (12 mentions) β actin, by Santa Cruz Biotechnology (9 mentions) Tnf α, by Abcam (8 mentions) Nlrp3, by Cell Signaling Technology (7 mentions) Ripa lysis buffer, by Beyotime (7 mentions) Bcl2 associated x (bax), by Abcam (6 mentions) β actin, by Cell Signaling Technology (6 mentions) Bca protein assay kit, by Thermo Fisher Scientific (6 mentions) Caspase 3, by Abcam (5 mentions) Caspase 11, by Abcam (5 mentions) Bcl 2, by Abcam (5 mentions) Caspase 3, by Cell Signaling Technology (5 mentions) Interleukin 6 (il 6), by Abcam (5 mentions)

Close spelling variants of this product

Primary antibodies against caspase-1 Caspase-1 active staining kit Anti-caspase-1 p10 Anti-human caspase-1 Rabbit anti-caspase-1 Caspase-1 activity assay kit Caspase-1 p10 Anti-mouse caspase-1 Anti-caspase-1 (p20; Caspase-1 Colorimetric Assay Kit

146 protocols using caspase 1

Western Blot Analysis of Apoptosis and Inflammation

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Mouse testis tissues or cells were lysed with RIPA lysis buffer. The protein concentrations were determined by a BCA protein detection kit (Beyotime, China). Western blotting was conducted following standard procedures. The antibodies used in this study were as follows: Caspase-3, Cleaved Caspase-3, Bax, Puma, Cytochrome-c, Parp, Cleaved Parp, p21, p27, Cdk2, Cdk4, CyclinD1, TGF-β, IL-1β, IL-18, Caspase-1, Gapdh (CST), TNF-α, Caspase-1, and Gsdmd antibodies (Abcam).

Ma J., Dong S., Lu H., Chen Z., Yu H., Sun X., Peng R., Li W., Wang S., Jiang Q., Li F, & Ma L. (2022). The hydrogen storage nanomaterial MgH2 improves irradiation-induced male fertility impairment by suppressing oxidative stress. Biomaterials Research, 26, 20.

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Western Blot Analysis of Inflammatory Markers

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Lysates were subjected to Western blot assay using anti-mouse IL-1β (R&D), caspase-1 (Abcam, Cambridge, MA) and NLRP3 (Cell Signaling) or anti-human IL-1β (R&D) or caspase-1 (Abcam).

Han M., Bentley J.K., Rajput C., Lei J., Ishikawa T., Jarman C.R., Lee J., Goldsmith A.M., Jackson W.T., Hoenerhoff M.J., Lewis T.C, & Hershenson M.B. (2019). Inflammasome activation is required for human rhinovirus-induced airway inflammation in naïve and allergen-sensitized mice. Mucosal immunology, 12(4), 958-968.

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Caspase Activities in Myocyte Lysates

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The Caspase-1 activity and caspase-3 activity in the myocyte lysates were determined using the Caspase-1 or Caspase-3 Colorimetric Assay Kit separately (BioVision, USA). According to instructions, a total of 5 × 10⁶ cells were collected, resuspended in ice-cold cell lyses buffer, incubated on ice for 10 min, and centrifuged at 10000×g for 1 min. The supernatant, which served as a detection sample, was collected and incubated with 2× reaction buffer (including 10 mM DTT) and 4 mM DEVD-pNA buffer for 2 h at 37 °C. A microplate reader was used to detect the absorbance at 405 nm. At least three independent experiments were performed.

Xiang F., Ma S.Y., Lv Y.L., Zhang D.X., Song H.P, & Huang Y.S. (2019). Tumor necrosis factor receptor-associated protein 1 regulates hypoxia-induced apoptosis through a mitochondria-dependent pathway mediated by cytochrome c oxidase subunit II. Burns & Trauma, 7, 16.

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Platelet Function and Signaling Assays

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Platelet receptor expression, activation, aggregation and immunoblotting were studied as previously described.^{17 (link),18 (link)} Antibodies against c-Src (anti-Tyr-416, Cell Signaling Technology; pan-c Src, Proteintech), Syk (anti-Tyr-525 and pan-Syk, Bioworld Technology) and PLCγ2 (anti-Tyr-1217 and pan-PLCγ2; Bioworld Technology), IL-1β (Cell Signaling Technology) and Caspase-1 (BioVision) were used.
Detailed methods of the electron microscopy of platelet spreading, and clot retraction are provided in the Online Supplement.

Qiao J., Wu X., Luo Q., Wei G., Xu M., Wu Y., Liu Y., Li X., Zi J., Ju W., Fu L., Chen C., Wu Q., Zhu S., Qi K., Li D., Li Z., Andrews R.K., Zeng L., Gardiner E.E, & Xu K. (2018). NLRP3 regulates platelet integrin αIIbβ3 outside-in signaling, hemostasis and arterial thrombosis. Haematologica, 103(9), 1568-1576.

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Evaluating Disaccharides' Effects on α-Synuclein Aggregation

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The SNCA-GFP SH-SY5Y cells were pretreated with each test disaccharide and induced SNCA-GFP expression in the presence of α-synuclein fibrils as described. For LDH release assay, cell culture media were collected on day 14 and the release of LDH was examined by using LDH cytotoxicity assay kit (Cayman, Ann Arbor, MI, USA). The absorbance was read at 490 nm with Multiskan GO microplate reader. For caspase 1 and 3 activity assays, cells were lysed in 1 × lysis buffer by repeated cycles of freezing and thawing. caspase 1 and 3 activities were measured using the caspase 1 (BioVision) and caspase 3 (Sigma-Aldrich) fluorimetric assay kits, with excitation/emission wavelengths of 420 ± 25/485 ± 10 nm (caspase 1 assay) or 360 ± 20/460 ± 20 nm (caspase 3 assay) (FLx800 fluorescence microplate reader, Bio-Tek).

Chen C.M., Lin C.H., Wu Y.R., Yen C.Y., Huang Y.T., Lin J.L., Lin C.Y., Chen W.L., Chao C.Y., Lee-Chen G.J., Su M.T, & Chang K.H. (2020). Lactulose and Melibiose Inhibit α-Synuclein Aggregation and Up-Regulate Autophagy to Reduce Neuronal Vulnerability. Cells, 9(5), 1230.

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Protein Antibody Detection in Lung Tissue

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Anti-NLRP3Ab, anti-Cas-1Ab, anti-GSDMDAb, anti-IL-1βAb, and anti-SP-B in lung tissue homogenate were detected by a commercially available, highly specific ELISA kits (NLRP3 kit, Aviva Systems Biology, San Diego, USA), (caspase-1, BioVision, Milpitas, USA), (GSDMD, Biomatik, Ontario, Canada), (IL-1β, Cloud-Clone Corp., Katy, USA), and (SP-B, Novus, Ontario, Canada). The microtiter plates coated with the specific protein to find the specific anti-protein Ab or with control proteins to find nonspecific interactions. The assay was performed according to the manufacturer’s instructions. Absorbance was measured using Labsystems Multiscan MS ELISA reader (DASIT).

El-Shaer N.O., Hegazy A.M, & Muhammad M.H. (2023). Protective effect of quercetin on pulmonary dysfunction in streptozotocin-induced diabetic rats via inhibition of NLRP3 signaling pathway. Environmental Science and Pollution Research International, 30(14), 42390-42398.

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Quantifying Apoptosis in A53T SNCA-GFP SH-SY5Y Cells

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TPA-differentiated A53T SNCA-GFP SH-SY5Y cells (4 × 10⁵ on 6-well dishes for caspase 1 activity and LDH release assays; 5 × 10⁴ on 12-well dishes for caspase 3 activity assay) were pretreated with test compounds or herbs and A53T SNCA-GFP expression was induced in the presence of α-synuclein fibrils as described. For LDH release assay, cell culture media were collected on day 14 and the release of LDH was examined by using LDH cytotoxicity assay kit (Cayman, Ann Arbor, MI, USA). The absorbance was read at 490 nm with Multiskan GO microplate reader. For caspase 1 and 3 activity assays, cells were lysed by repeated cycles of freezing and thawing. caspase 1/3 activities were measured with the caspase 1 (BioVision, Milpitas, CA, USA) and caspase 3 (Sigma-Aldrich) fluorimetric assay kits, with 420/50 nm excitation and 485/20 nm emission (caspase 1 assay) or 360/40 nm excitation and 460/40 nm emission (caspase 3 assay) (FLx800 fluorescence microplate reader, Bio-Tek).

Chen C.M., Yen C.Y., Chen W.L., Lin C.H., Wu Y.R., Chang K.H, & Lee-Chen G.J. (2021). Pathomechanism Characterization and Potential Therapeutics Identification for Parkinson’s Disease Targeting Neuroinflammation. International Journal of Molecular Sciences, 22(3), 1062.

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Neuroblastoma Cell Viability Assay

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SK-N-LP/luciferase cells were seeded in a 96-well plate (around 4000 cells/well). After overnight culture, SK-N-LP/luciferase cells were pre-treated with DMSO, TNFα antibody (Sigma-Aldrich, St. Louis, USA), caspase 1 (Biovision, San Diego, USA), and pan-caspase (Calbiochem, Darmstad, Germany) inhibitors for 2 h, respectively. The viability of neuroblastoma with Salmonella or macrophages supernatant treatment was observed by XTT kit (Roche, Mannheim, Germany) for three continuous days or after 2 days, respectively. The optical density was measured using a microplate reader at wavelength 450 nm.

Guo Z.L., Yu B., Ning B.T., Chan S., Lin Q.B., Li J.C., Huang J.D, & Chan G.C. (2015). Genetically modified "obligate" anaerobic Salmonella typhimurium as a therapeutic strategy for neuroblastoma. Journal of Hematology & Oncology, 8, 99.

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Microinjection-based GSDMB and GSDMD Activation

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H1299 cells (WT or stably expressing GSDMB-Flag or GSDMD) plated at 150,000 cells/well in 60 mm culture dishes 2 days before microinjection. Just prior to microinjection, culture media was replaced with media supplemented with 20 mM HEPES. Cells were located using an Axio Observer.A1 microscope (Zeiss) and microinjected using pulled glass pipettes (Sutter Instruments P-97) and an Eppendorf FemtoJet and InjectMan NI 2 according to manufacturer’s instructions. Cells were injected with a solution containing 2 mg/ml purified GZMA (WT and GSDMB cells) or 0.125 U/μl Caspase 1 (Biovision 1081-25; WT and GSDMD cells) along with 1 mg/ml of either Texas Red Dextran 10,000 MW (Invitrogen D1863 ) or Cascade Blue Dextran 10,000 MW (Invitrogen D1976) in PBS. Live microinjected cells were identified by Texas Red fluorescence and the same field of live cells was imaged at 0h, 2h and 4 h after microinjection using an Eclipse Ti-S microscope (Nikon). Cells were then washed 3 times with PBS, fixed in 4% paraformaldehyde (in PBS) for 10 min., washed 3 times in PBS and coverslips were mounted onto glass slides using VectaShield Plus Antifade Mounting Medium (VectaShield H1900). Microinjected cells were identified by Cascade blue labeling and imaged on an Observer.Z1 inverted microscope (Zeiss).

Hansen J.M., de Jong M.F., Wu Q., Zhang L.S., Heisler D.B., Alto L.T, & Alto N.M. (2021). Pathogenic Ubiquitination of GSDMB Inhibits NK-cell Bactericidal Functions.. Cell, 184(12), 3178-3191.e18.

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Retinal Protein Extraction and Western Blot

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Proteins were extracted from mice retinas by sonication in lysis buffer RIPA buffer (pH = 8) containing 50 mM Tris-HCl, 150 mM NaCl, 5 mM EDTA, 1% Triton 100×, 0.5% sodium deoxycholate, 0.1% SDS, and a cocktail of protease and phosphatase inhibitors (MiniComplete, PhosphoStop, and PMSF, Roche, Bâle, Switzerland). Protein concentrations were determined using the Bicinchoninic Acid Protein Assay Kit (Pierce, Rockford, IL, USA). Thirty micrograms of protein per sample were electrophoresed on 15% sodium dodecyl sulfate-polyacrylamide gels using an electrophoresis system (Mini-Protean Tetra System, Bio-Rad, Hercules, CA, USA) and then transferred onto polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). Membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween-20 for 1 h at room temperature, and incubated with the following primary antibodies: 1:800 IL-1β (Abcam, ab9722), 1:500 caspase-1 (BioVision, 3019), and 1:1000 β-actin (Santa Cruz, sc47778) then incubated with 1:6000 horseradish peroxidase-conjugated secondary antibodies (Millipore, AP307P and Millipore, AP308P). Densitometric analysis of western blotting bands was quantified using the ImageJ software and normalized to β-actin.

Dabouz R., Cheng C.W., Abram P., Omri S., Cagnone G., Sawmy K.V., Joyal J.S., Desjarlais M., Olson D., Weil A.G., Lubell W., Rivera J.C, & Chemtob S. (2020). An allosteric interleukin-1 receptor modulator mitigates inflammation and photoreceptor toxicity in a model of retinal degeneration. Journal of Neuroinflammation, 17, 359.

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