Log In Sign up
The largest database of trusted experimental protocols
> Chemicals & Drugs > Amino Acid > Procaspase-1

Procaspase-1

Procaspase-1 is an inactive precursor of the protease caspase-1, which plays a crucial role in the inflammatory response.
It is involved in the proteolytic activation of proinflammatory cytokines, such as interleukin-1β and interleukin-18.
Researching Procaspase-1 can provide important insights into the mechanisms of inflammation and offer potential therapeutic targets for inflammatory diseases.
PubCompare.ai's AI-driven protocol comparison tool can optimize your Procaspase-1 research by helping you easily locate and identify the best experimental protocols from literature, preprints, and patents.
This can ensure reproducibility and accuracy in your experiments, while also leveraging AI-powered analysis to compare protocols side-by-side and make informed decisions about your experimental setup.
Improve the quality and efficieny of your Procaspase-1 research with PubCompare.ai.

Most cited protocols related to «Procaspase-1»

1
Recombinant Expression of Inflammasome Proteins
Cited 4 times
cDNAs for mouse Nlrp3, pro-Casp1, pro-IL1β, ASC and Gsdmd were amplified from reverse-transcribed cDNAs of J774 cells. Nlrp3, pro-casp1, pro-IL1β, ASC and Gsdmd cDNAs were inserted into a modified pBOBi plasmid vector with N-terminal or C-terminal 3× Flag or 3× HA tag. Truncation or point mutation of Gsdmd was generated by standard PCR cloning strategy. All plasmids were DNA-sequenced.
Antibody for IL-1β (5129-100) was purchased from Biovision. Antibody for Caspase-1 p20 (clone 4B4) was a kind gift from VM Dixit (Genetech, USA). Antibody for NLRP3 (AG-20B-0006-C100) was purchased from Adipogen. Antibody for GSDMD was purchased from Proteintech. Mouse anti-HA (F-7), anti-GAPDH (6C5) and ASC (sc-22514-R) antibodies were purchased from Santa Cruz Biotechnology, Inc. Mouse anti-Flag M2 and anti-HA beads, and anti-β-Tubulin (T6199) antibodies were purchased from Sigma-Aldrich. Ultrapure LPS from E. coli O111:B4, PI and Cholera Toxin B subunit were purchased from Sigma-Aldrich. Nigericin was purchased from InvivoGen. zVAD was obtained from Calbiochem. Anthrax Lethal Factor and Protective Antigen were purchased from List Biological Laboratories, Inc. CytoTox 96 Non-Radioactive Cytotoxicity Assay kit was purchased from Promega.
He W.T., Wan H., Hu L., Chen P., Wang X., Huang Z., Yang Z.H., Zhong C.Q, & Han J. (2015). Gasdermin D is an executor of pyroptosis and required for interleukin-1β secretion. Cell Research, 25(12), 1285-1298.
Publication 2015
anthrax lethal factor Antibodies Antigens Biological Assay Biopharmaceuticals Caspase 1 Cells Choleragenoid Clone Cells Cloning Vectors Cytotoxin DNA, Complementary Escherichia coli GAPDH protein, human Immunoglobulins Interleukin-1 beta Mus Nigericin Nod-like receptor protein 3 inflammasome, mouse Plasmids Point Mutation Procaspase-1 Promega Radioactivity Tubulin
2
Bladder Epithelial Cell Gene Expression
Cited 3 times
Total RNA was isolated from bladder epithelial cells using E.Z.N.A. ® Total RNA Kit I (Omega Bio-tek, GA, USA) according to manufacturer's protocol. The concentration of RNA was determined using spectrophotometry (Nano- Drop ND-1000, Wilmington, NC, USA). First strand cDNA synthesis was performed by High capacity cDNA RT kit (Thermo Fisher Scientific). The real time-RT-PCR was carried out in a total volume of 10 μl on a 96-well plate (Sarstedt, Nümbrecht, Germany). Each well contained 5 μl Maxima SYBR Green qPCR Master Mix (Thermofisher), 1 μl of each primer IL-1β (Forward: 5′-CCACAGACCTTCCAGGAGAATG-3, Revers: 5′-GTGCAGTTCAGTGATCGTACAGG-3′), NLRP3 (Forward: 5′-GGACTGAAGCACCTGTTGTGCA-3, Revers: 5′-TCCTGAGTCTCCCAAGGCATTC-3′), pro-caspase-1 (Forward: 5′-GCTGAGGTTGACATCACAGGCA-3, Revers: 5′-TGCTGTCAGAGGTCTTGTGCTC-3′), ASC (Forward: 5′-AGCTCACCGCTAACGTGCTGC-3, Revers: 5′-GCTTGGCTGCCGACTGAGGAG-3′), uroplakin 1a (Forward: 5′-CACCAAGCAGATGCTGACCTTC−3, Revers: 5′-GGACCAGATGTGCCACAGCATT-3′), uroplakin 3a (Forward: 5′-CTCACAGATCCTGAATGCCTACC-3, Revers: 5′-CCGTGGACATATTGACCAGGAC-3′), integrin α3 (Forward: 5′-GCCTGACAACAAGTGTGAGAGC-3, Revers: 5′-GGTGTTCGTCACGTTGATGCTC-3′), integrin β1 (Forward: 5′-GGATTCTCCAGAAGGTGGTTTCG-3, Revers: 5′-TGCCACCAAGTTTCCCATCTCC-3′), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Forward: 5′-GTCTCCTCTGACTTCAACAGCG-3, Revers: 5′-ACCACCCTGTTGCTGTAGCCAA-3′) (Eurofins MWG Synthesis GmbH, Ebersberg, Munich), 1 μl cDNA (10 ng) and 3 μl RNase free water. The PCR amplification was conducted in a CFX96 Touch™ Real-Time PCR Detection System (Biorad, Hercules, CA, USA) using the following protocol: initial denaturation at 95°C for 10 min, 40 cycles of denaturation at 95°C for 15 s followed by annealing/extension at 60°C for 60 s. Each PCR was followed by a dissociation curve analysis between 60-95°C. The Ct values were analyzed by the comparative Ct (ΔΔCt) method and normalized to the endogenous control GAPDH. Fold difference was calculated as 2−ΔΔCt.
Demirel I., Persson A., Brauner A., Särndahl E., Kruse R, & Persson K. (2018). Activation of the NLRP3 Inflammasome Pathway by Uropathogenic Escherichia coli Is Virulence Factor-Dependent and Influences Colonization of Bladder Epithelial Cells. Frontiers in Cellular and Infection Microbiology, 8, 81.
Full text: Click here
Publication 2018
Anabolism DNA, Complementary Endoribonucleases Epithelial Cells Glyceraldehyde-3-Phosphate Dehydrogenases Integrins Interleukin-1 beta Oligonucleotide Primers Procaspase-1 Real-Time Polymerase Chain Reaction RNA I Spectrophotometry SYBR Green I Touch Urinary Bladder Uroplakin Ia Uroplakins
3
NR1D1 Regulates NLRP3-Mediated Cytokine Secretion
Cited 3 times

Protocol full text hidden due to copyright restrictions

Open the protocol to access the free full text link

Publication 2017
Caspase Caspase-8 Caspase 1 Culture Media Cytokinesis Deletion Mutation Gene Expression Hemin Homo sapiens IL1B protein, human Inflammasomes Interleukin-18 Ligands Macrophage MCC-950 methylene dimethanesulfonate Procaspase-1 Proteins Protein Subunits Psychological Inhibition secretion Transcription, Genetic
4
Reconstituted HEK293 Inflammasome Assay
Cited 3 times

Protocol full text hidden due to copyright restrictions

Open the protocol to access the free full text link

Publication 2012
Cells HEK293 Cells Homo sapiens Interleukin-1 beta lipofectamine 2000 Mus Procaspase-1 Transfection
5
Molecular Cloning of ASC Isoforms
Cited 3 times
A search of the publicly available expressed sequence tag (EST) database revealed three potential ASC isoforms: ASC-b (Acc. No. BM456838), ASC-c (Acc. No. BE560228), and ASC-d (Acc. No. BM920038). The complete open reading frame of each isoform was subsequently amplified by PCR from pooled THP-1 cell cDNAs that were induced with a cocktail of cytokines for 2 to 24 hours. ASC-b, ASC-c, and ASC-d were amplified using the common forward primer 5'-CGGAATTCGATCCTGGAGCCATGGG-3' and the common reverse primer 5'-CGCTCGAGTGACCGGAGTGTTGCTG-3' and cloned into a modified pcDNA3 vector (Invitrogen) in frame with an NH2-terminal myc epitope tag. The CARD of caspase 1 was amplified by high fidelity PCR and cloned into pGex4T-1 (Novagen). All other expression constructs (ASC, pro-IL-1β, pro-caspase 1, NLRP3R260W) have been previously described [37 (link)-39 (link)].
Bryan N.B., Dorfleutner A., Kramer S.J., Yun C., Rojanasakul Y, & Stehlik C. (2010). Differential splicing of the apoptosis-associated speck like protein containing a caspase recruitment domain (ASC) regulates inflammasomes. Journal of Inflammation (London, England), 7, 23.
Full text: Click here
Publication 2010
Caspase 1 Cloning Vectors Cytokine DNA, Complementary Epitopes Interleukin-1 beta Oligonucleotide Primers Procaspase-1 Protein Isoforms Reading Frames THP-1 Cells

Most recents protocols related to «Procaspase-1»

1
Protein Extraction and Immunoblotting Protocol
Protein samples from supernatants and cell lysates were prepared as previously described [25 (link)]. The membrane transfer parameters for pro-IL-1β, pro-caspase-1, caspase-1 p20, NLRP3, β-actin and His-tag were 200 mA/90 min. The primary antibody dilution rates were 1:1000 for targets of interleukin-1β (IL-1β; R&D Systems, Minneapolis, MN, USA), caspase-1 (p20) (Adipogen, Switzerland), and NLRP3 (Adipogen SA, Epalinges, Switzerland) and 1:5000 for targets of His-tag (Amylet Scientific, Wuhan, China) and β-actin (Proteintech, Wuhan, China).
Zhao P., Li J., Li X., Dong J., Wang X., Zhang N., Li S., Sun M., Zhang X., Wang Z., Liang M., Li Y., Cao L, & Gong P. (2023). The NLRP3 inflammasome recognizes alpha-2 and alpha-7.3 giardins and decreases the pathogenicity of Giardia duodenalis in mice. Parasites & Vectors, 16, 85.
Full text: Click here
Publication 2023
Actins Caspase 1 Cells IL1B protein, human Immunoglobulins Interleukin-1 beta Procaspase-1 Proteins Technique, Dilution Tissue, Membrane
2
Western Blot Analysis of Apoptosis Markers
Protein extracts were separated on 8–15% polyacrylamide gels and then transferred onto polyvinylidene difluoride (PVDF) membranes (MilliporeSigma, Burlington, MA, USA). After blocking nonspecific binding with 5% nonfat dry milk (Sangon Biotech Shanghai Co., Ltd.) membranes were probed with primary antibodies overnight at 4°C and incubated with ppropriate HRP-labeled secondary antibodies at room temperature for 1 h. Membranes were then visualized with an enhanced chemiluminescence luminescence reagent (Meilunbio). The intensities of the bands were determined by ImageJ Launcher broken symmetry software program (NIH, Bethesda, MD, USA). Following antibodies were used: rabbit anti-mouse cleaved caspase-8 (#8592; Cell Signaling Technology), rabbit anti caspase-8 (AC056; Beyotime), Phospho-RSK2 (Ser227) Rabbit mAb (#3556S; Cell Signaling Technology), PDK1 (D37A7) Rabbit mAb (#5662S; Cell Signaling Technology), Caspase-3 (D3R6Y) Rabbit mAb (#14220S; Cell Signaling Technology), Caspase-1 (D7F10) Rabbit mAb (#3866S; Cell Signaling Technology), Anti-MLKL (phospho S345) (ab196436; Abcam), Anti-MLKL (phospho S358) (ab187091; Abcam), Anti-pro Caspase-1 + p10 + p12 (ab179515; Abcam), Anti-GSDMD antibody (ab209845; Abcam), Anti-GSDMD antibody (ab225867; Abcam), Anti-cleaved N-terminal GSDMD (ab215203; Abcam), anti-beta-actin (bs-0061 R; Bioss).
Yuan H., Zhou L., Chen Y., You J., Hu H., Li Y., Huang R, & Wu S. (2023). Salmonella effector SopF regulates PANoptosis of intestinal epithelial cells to aggravate systemic infection. Gut Microbes, 15(1), 2180315.
Full text: Click here
Publication 2023
Antibodies Antibodies, Anti-Idiotypic beta-Actin Caspase-8 Caspase 1 Caspase 3 Chemiluminescence Luminescence Milk, Cow's Mus PDK1 protein, human polyacrylamide gels polyvinylidene fluoride Procaspase-1 Proteins Rabbits RPS6KA3 protein, human Tissue, Membrane
3
Western Blot Analysis of NLRP3 Inflammasome
Tissue and cellular samples were homogenized in RIPA lysis buffer (Beyotime, Shanghai, China) with phosphatase inhibitor cocktail and protease inhibitors (MedChemExpress). Protein concentration of extracts of PMVECs or lung tissue homogenates was examine using a BCA protein assay kit (Beyotime). Protein samples (15 μg) were loaded on sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and transferred onto a polyvinylidene fluoride (PVDF) membrane. After blocking with 5% skimmed milk for 1 h, the membrane was incubated overnight with primary antibodies against ASC (ab180799, 1:1000; Abcam), caspase‐1 p20 (WL02996a, 1:1000; Wanleibio, Shenyang, China), MyD88 (ab219413, 1:1000; Abcam), procaspase‐1 (ab179515, 1:1000; Abcam), GSDMD‐N (#397545, 1:1000; Cell signaling Technology, Shanghai, China), NLRP3 (ab263899, 1:1000; Abcam), pro‐GSDMD (#464515, 1:1000; Cell signaling), GAPDH (ab181602, 1:10000; Abcam), TLR4 (sc‐293 072, 1:1000; San Cruz Biotechnology, Dallas, TX, USA), p‐p65 (ab76302, 1:1000; Abcam), and p65 (ab19870, 2.5 μg/ml; Abcam) at 4 °C. The membrane was then washed and incubated with secondary antibodies (1:20000) for 1 h at room temperature. An enhanced chemiluminescence detection system (Yeasen, Shanghai, China) was used to detect antibody‐specific proteins. Band densitometry analysis was performed using ImageJ software.
Jin T., Ai F., Zhou J., Kong L., Xiong Z., Wang D., Lu R., Chen Z, & Zhang M. (2023). Emodin alleviates lung ischemia–reperfusion injury by suppressing gasdermin D‐mediated pyroptosis in rats. The Clinical Respiratory Journal, 17(3), 241-250.
Full text: Click here
Publication 2023
Antibodies Antibody Specificity Biological Assay Buffers Caspase 1 Cells Chemiluminescence Densitometry GAPDH protein, human Lung Milk, Cow's Phosphoric Monoester Hydrolases polyvinylidene fluoride Procaspase-1 Protease Inhibitors Proteins Radioimmunoprecipitation Assay SDS-PAGE Tissue, Membrane Tissues
4
Immunoblotting for Cell Signaling
Cells were lysed in pre-cooled RIPA lysis buffer (Beyotime) with freshly added protease inhibitor mixture (Hoffman-La Roche Ltd., Basel, Switzerland). The protein concentration was determined by the Dioctanoic acid determination kit (Thermo Fisher Scientific). The same amount of protein was separated by SDS-PAGE and transferred to PVDF membrane. The membrane and the first antibody (Bcl-2, Bax, pro-caspase-1, NLRP3, GAPDH) were incubated at 4°C overnight. After incubation with HRP-labeled secondary antibody at room temperature for 2 h, the protein bands were analyzed using ECL Hypersensitive Chemiluminescence Kit (Beyotime).
Wei S., Chen Z., Ling X., Zhang W, & Jiang L. (2023). Comprehensive analysis illustrating the role of PANoptosis-related genes in lung cancer based on bioinformatic algorithms and experiments. Frontiers in Pharmacology, 14, 1115221.
Full text: Click here
Publication 2023
Acids BCL2 protein, human Buffers Cells Chemiluminescence GAPDH protein, human Hypersensitivity Immunoglobulins polyvinylidene fluoride Procaspase-1 Protease Inhibitors Proteins Radioimmunoprecipitation Assay SDS-PAGE Tissue, Membrane
5
NLRP3 Inflammasome Pathway Activation
Cell lysates were collected by centrifugation (10,000× g) at 4 °C for 30 min. After centrifugation, the proteins in the supernatant were subjected to separation by sodium dodecylsulfate polyacrylamide gel electrophoresis, and transferred onto a polyvinylidene difluoride membrane. The membrane was immersed in 5% non-fat milk for 1 h prior to incubation with the primary antibodies at 4 °C overnight. Primary antibodies against GAPDH (1:5000), NLRP3 (1:1000), caspase-1 (1:1000), pro-caspase-1 (1:1000), ASC (1:1000), p-NF-κB (1:1000), NF-κB (1:1000), IL-1β (1:1000), p-ERK1/2 (1:1000), ERK1/2 (1:1000), p-JNK (1:1000), and JNK (1:1000) were used in the immunological detection. Membranes were rinsed with Tris-buffered saline containing 0.1% Tween® 20 (TBST) 3 times prior to incubation with secondary antibodies at 37 °C for 1 h. Then, the target protein bands were imaged using the enhanced chemiluminescent reagent (Invitrogen) and quantitated by the iBright Imaging Systems (iBright FL 1000; Thermo Fisher Scientific, Inc., Waltham, MA, USA).
Huang K.C., Chang Y.T., Pranata R., Cheng Y.H., Chen Y.C., Kuo P.C., Huang Y.H., Tzen J.T, & Chen R.J. (2023). Alleviation of Hyperuricemia by Strictinin in AML12 Mouse Hepatocytes Treated with Xanthine and in Mice Treated with Potassium Oxonate. Biology, 12(2), 329.
Full text: Click here
Publication 2023
Antibodies Caspase 1 Cells Centrifugation GAPDH protein, human Interleukin-1 beta Milk, Cow's Mitogen-Activated Protein Kinase 3 polyvinylidene fluoride Procaspase-1 Proteins RELA protein, human Saline Solution SDS-PAGE Tissue, Membrane Tween 20

Top products related to «Procaspase-1»

1
Ab179515 by Abcam
Sourced in United States, United Kingdom, China
Ab179515 is a laboratory reagent produced by Abcam. It is a monoclonal antibody that can be used for research applications. The core function of this product is to detect and bind to a specific target molecule.
2
Pvdf membrane by Merck Group
Sourced in United States, Germany, China, United Kingdom, Morocco, Ireland, France, Italy, Japan, Canada, Spain, Switzerland, New Zealand, India, Hong Kong, Sao Tome and Principe, Sweden, Netherlands, Australia, Belgium, Austria
PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
3
Pro caspase 1 by Abcam
Sourced in United States, United Kingdom
Pro-caspase-1 is a protein involved in the activation of the inflammatory caspase-1 enzyme. Caspase-1 plays a key role in the processing and maturation of the pro-inflammatory cytokines IL-1β and IL-18. Pro-caspase-1 is the inactive precursor form of caspase-1 that requires cleavage for activation.
4
Ripa lysis buffer by Beyotime
Sourced in China, United States, Switzerland, Germany, Japan, United Kingdom
RIPA lysis buffer is a detergent-based buffer solution designed for the extraction and solubilization of proteins from cells and tissues. It contains a mixture of ionic and non-ionic detergents that disrupt cell membranes and solubilize cellular proteins. The buffer also includes additional components that help to maintain the stability and activity of the extracted proteins.
5
Gapdh by Cell Signaling Technology
Sourced in United States, United Kingdom, China, Germany, Japan, Canada, Morocco, France, Netherlands, Ireland, Israel, Australia, Sweden, Macao, Switzerland
GAPDH is a protein that functions as an enzyme involved in the glycolysis process, catalyzing the conversion of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate. It is a common reference or housekeeping protein used in various assays and analyses.
6
Pro caspase 1 by Santa Cruz Biotechnology
Sourced in United States
Pro-caspase-1 is a recombinant human pro-caspase-1 protein. Caspase-1 is a protease enzyme involved in the inflammatory response and programmed cell death. The pro-form of the enzyme requires activation to become catalytically active.
7
Il 1β by Abcam
Sourced in United States, United Kingdom, China, Canada
IL-1β is a recombinant human interleukin-1 beta protein. Interleukin-1 beta is a pro-inflammatory cytokine that plays a central role in the regulation of immune and inflammatory responses.
8
Il 1β by Cell Signaling Technology
Sourced in United States, United Kingdom, China, Germany
IL-1β is a cytokine that plays a crucial role in the inflammatory response. It functions as a mediator of the innate immune system.
9
Pro caspase 1 by Cell Signaling Technology
Sourced in United States
Pro-caspase-1 is a laboratory reagent used for research purposes. It is the inactive precursor form of the caspase-1 enzyme, which plays a role in the regulation of inflammation and cell death. The product is intended for use in biochemical and cell-based assays to study caspase-1 activation and related cellular processes.
10
Bca protein assay kit by Beyotime
Sourced in China, United States, Germany, Puerto Rico, United Kingdom, Switzerland, Japan, Sweden
The BCA protein assay kit is a colorimetric-based method for the quantitative determination of total protein concentration in a sample. It uses bicinchoninic acid (BCA) to detect and quantify the presence of protein.

More about "Procaspase-1"

Procaspase-1, also known as Caspase-1 Precursor or Pro-caspase-1, is an inactive form of the protease enzyme caspase-1.
Caspase-1 plays a crucial role in the inflammatory response by proteolytically activating pro-inflammatory cytokines such as interleukin-1β (IL-1β) and interleukin-18 (IL-18).
Researching Procaspase-1 can provide valuable insights into the mechanisms of inflammation, which is essential for understanding and developing potential therapeutic targets for inflammatory diseases.
To study Procaspase-1, researchers often utilize various experimental techniques and protocols.
One common method is Western blotting, where Procaspase-1 levels can be detected using specific antibodies, such as Ab179515.
PVDF membranes are frequently used for protein transfer during Western blotting.
Additionally, RIPA lysis buffer is commonly employed to extract and solubilize Procaspase-1 from cellular samples.
The expression of Procaspase-1 is often normalized against a housekeeping protein, such as GAPDH, to ensure accurate quantification.
The activation of Procaspase-1 leads to the cleavage and release of mature IL-1β, a potent pro-inflammatory cytokine.
Measuring IL-1β levels can provide valuable insights into Procaspase-1 activity and the inflammatory response.
Techniques like ELISA or BCA protein assay kits can be used to quantify IL-1β and total protein levels, respectively.
PubCompare.ai's AI-driven protocol comparison tool can optimize your Procaspase-1 research by helping you easily locate and identify the best experimental protocols from literature, preprints, and patents.
This can ensure reproducibility and accuracy in your experiments, while also leveraging AI-powered analysis to compare protocols side-by-side and make informed decisions about your experimental setup.
Improve the qulaity and efficeiny of your Procaspase-1 research with PubCompare.ai.