Commercially available antibodies were obtained as follows: anti-caspase 12, anti-caspase 3, anti-caspase 9, anti-cleaved caspase 1, anti-p-JNK, anti-JNK, anti-p-eIF2α, anti-eIF2α, anti-IRE1α, anti-CHOP, anti-NLRP3, anti-COX IV, anti-IL1β, anti-cleaved caspase1, anti-p-IκB, anti-IκB, anti-NFκB, and anti-cytochrome c antibodies were from Cell Signaling Technology; anti-ATF6 and anti-p-IRE1α antibodies were from Novus Biological; anti-p-ASK1(Ser967) antibody was from Sigma-Aldrich; anti-XBP1s antibody was from BioLegend; anti-TXNIP antibody was from MBL International Corporation; anti-WT-1 antibodies were from Santa Cruz Biotechnology or Abcam; and anti-ATF4 and anti-Trx2 antibodies were from Santa Cruz Biotechnology. HRP-conjugated anti-mouse β-actin antibody was from Sigma-Aldrich and HRP-conjugated secondary antibodies were from Cell Signaling Technology.
Anti cleaved caspase 1
Manufactured by Cell Signaling Technology
Sourced in United States, China
Anti-cleaved caspase-1 is a laboratory product provided by Cell Signaling Technology. It is an antibody that specifically recognizes the cleaved form of the caspase-1 protein.
Automatically generated - may contain errors
Lab products found in correlation
Anti nlrp3, by Cell Signaling Technology (8 mentions)
Anti cleaved il 1β, by Cell Signaling Technology (7 mentions)
Anti caspase 1, by Cell Signaling Technology (5 mentions)
Anti β actin, by Cell Signaling Technology (5 mentions)
Anti pro caspase 1, by Cell Signaling Technology (3 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (3 mentions)
Anti cox 4, by Cell Signaling Technology (2 mentions)
Anti chop, by Cell Signaling Technology (2 mentions)
Anti il 1β, by Cell Signaling Technology (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Anti txnip, by Cell Signaling Technology (2 mentions)
Ly294002, by Merck Group (2 mentions)
Anti p akt, by Cell Signaling Technology (2 mentions)
Anti t akt, by Cell Signaling Technology (2 mentions)
Anti il 1β, by Santa Cruz Biotechnology (2 mentions)
Anti gapdh, by Bioworld Technology (2 mentions)
Anti caspase 1, by Thermo Fisher Scientific (2 mentions)
Anti nlrp3, by Abcam (2 mentions)
Anti β actin, by Abcam (2 mentions)
Hrp conjugated secondary antibody, by Cell Signaling Technology (1 mentions)
27 protocols using anti cleaved caspase 1
1
Comprehensive Antibody Characterization Protocol
2
Western Blotting for Inflammasome Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All western blotting procedures were performed as previously described (30 (link)) with the following primary antibodies: anti-PRX3, anti-NLRP3 (Abclonal Biotechnology, Wuhan, China), anti-gasdermin D (GSDMD), anti-caspase-1, anti-cleaved caspase-1, anti-IL-1β (all from Cell Signaling Technology, Boston, MA, USA), anti-IL-18, anti-PRX5 (Abcam, Cambridge, UK), PRX6 (Zen bio., Chengdu, China) and anti-β-actin (Bimake, Houston, TX, USA).
3
Western Blot Analysis of Protein Fractions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein samples were harvested from H9c2 cardiomyocytes after H/R and the myocardium at 24 hours after resuscitation. The separation of cytosolic and nuclear fractions was performed by centrifugation of samples at 16 000g. Subsequently, the samples were separated by SDS‐PAGE, then transferred to a polyvinylidene fluoride membrane, and finally blocked with 5% nonfat milk. After that, the membranes were incubated with primary anti‐NLRP3 (1:1000, Proteintech, Rosemount, IL), anticleaved caspase‐1 (1:1000, Cell Signaling Technology Inc., Danvers, MA), anti–gasdermin D (1:1000, Proteintech), anti‐HDAC6 (1:1000, Proteintech), anti‐GAPDH (1:5000, BBI Life Science Corporation, Shanghai, China), antiacetylated α‐tubulin (1:1000, Proteintech), anti–α‐tubulin (1:1000, Proteintech), anti‐TFEB (1:1000, Proteintech), antiacetylated lysine (1:1000, Cell Signaling Technology Inc.), antihistone H3 (1:1000, Abcam) at 4 °C for 24 hours, then rinsed with TBST solution, and finally incubated with the secondary antibody (1:5000, BBI Life Science Corporation) at room temperature for 1 hour. The protein was visualized with enhanced chemiluminescence substrates and analyzed by Image J software (National Institutes of Health, Bethesda, MD).
4
Immunofluorescence Staining of Liver Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunofluorescence was performed on frozen liver sections, as previously described38 (link). Liver sections were stained with the following primary antibodies: FITC Mouse Anti-iNOS/NOS Type II (1:200; BD Transduction Laboratories™, San Jose, CA, USA), PE anti-mouse CD206 (MMR) (1:200; BioLegend Inc., San Diego, CA, USA), Anti-actin, α-Smooth Muscle antibody (1:300; Sigma, St Louis, MO, USA), Anti-mouse MLKL (1:400; Biorbyt, San Francisco, CA, USA), Anti-MLKL (phospho S345) antibody (1:400; Abcam), Anti-RIPK3 (1:400; Abcam), RIP(D94C12) XP Rabbit mAb (1:400; Cell Signaling Technology, MA, USA), anti-NLRP3 antibody (1:300; Abcam), anti-ASC antibody (1:200, Cell Signaling Technology), and anti-cleaved caspase-1 (1:200; Cell Signaling Technology). For indirect immunofluorescent staining, liver sections were incubated with the following secondary antibodies: PE-conjugated donkey anti-rabbit IgG for p-MLKL, MLKL, RIPK3, and SMA (1:500); Alexa Fluor 488-conjugated donkey anti-rabbit IgG for p-MLKL, MLKL, RIPK3, RIPK1, NLRP3, ASC, and cleaved caspase-1 (1:500). Nikon Inverted Fluorescence Microscope ECLIPSE Ti and NIS-Elements F 3.0 Software (Nikon Corporation, Tokyo, Japan) were applied for image capture.
5
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The brain tissues of mice and cells were homogenized in lysis buffer containing protease inhibitors. The homogenate was centrifuged at 14000 g for 15 min at 4°C and the protein concentration was determined using the BCA kit. 30 μg lysate was loaded onto 10% SDS-PAGE. The proteins were transferred to PVDF membranes (Millipore, MA, USA). The membranes were blocked for 1 h in 5% dry milk and then incubated overnight with one of the following primary antibodies: anti-iNOS (ab178945), anti-Pro-caspase-1 (ab179515), anti-ANT (ab102032), anti-Cyp D (ab16045) (Abcam, Cambridge, MA, USA), anti-TH (#2792), anti-COX2 (#12282), anti-p-p65 (#3033), anti-Cleaved-caspase-1 (#89332), anti-Cyto C (#4280), anti-VDAC (#4866), anti-COX4 (#4850), or anti-β-actin (#3700) (Cell Signaling Technology, Beverly, USA). After washing 3 times in TBST for 5 min each, the membranes were incubated with goat anti-mouse, anti-rabbit, or anti-rat HRP for 1 h at room temperature. Then, the membranes were washed 3 times in TBST for 5 min each. The signal was visualized using an ECL chemiluminescence kit (Amersham Biosciences/GE Healthcare; Piscataway, NJ).
6
Inflammasome Activation Pathway Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Anti-IL-1β, Anti-Caspase1, TNF-α were purchased from R&D Systems (Minneapolis, MN, USA); Anti-cleaved caspase1, Anti-NLRP3, Anti-IκBα, Anti-phospho-P38, Anti-phospho-ERK, Anti-phospho-JNK, Anti-phospho-IKKα/β, and Anti-Tublin were purchased from Cell Signaling Technology (Boston, MA, USA); Anti-TAK1, Anti-phospho-TAK1, Anti-ASC and recombinant active caspase1 protein were purchased from Abcam (Shanghai, China). The ultrapure LPS and Nigericin were obtained from Sigma-Aldrich (St. Louis, MO, USA). The caspase1 activity assay kit was purchased from Beyotime Biotechnology (Shanghai, China). The Human IL-1β ELISA Kit was purchased from Boster Biological Technology (Wuhan, China).
7
Multiparametric Immunofluorescence Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
As previously described [18 (link)], frozen sections were fixed with 4% paraformaldehyde, washed with phosphate buffer saline (PBS), permeabilized with 0.2% Triton X-100, and blocked with 5% FBS, followed by incubation with primary antibodies anti-CD68 (Cat# ab53444, Abcam), anti-smooth muscle cell (SMC) α-actin (Cat# ab21027, Abcam), anti-cleaved caspase-1 (Cat# 3866, Cell Signaling Technology), anti-cleaved IL-1β (Cat# AB41610, Absci), and anti-Nur77 (Cat# ab153914, Abcam) overnight. The samples were then incubated with secondary antibodies and counterstained with 4′,6-diamidino-2-phenylindole. Fluorescent images were finally taken with a Zeiss LSM laser confocal scanning microscope within 24 hours.
8
Western Blot Analysis of Protein Targets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein lysate were prepared in RIPA lysis buffer (Pierce, Rockford, USA). 30 μg of total proteins were separated by SDS-PAGE, and transferred onto PVDF membrane (Pierce). After blocking with 5% non-fat milk, the blots were incubated with primary antibody at 4 °C overnight. The blots were incubated with HRP-conjugated secondary antibody (Invitrogen). The signal was detected using ECL substrate (Pierce). The following antibodies were used in this study: anti-TXNIP (#14715; 1:1000), anti-NLRP3 (#15101; 1:1000), anti-ASC (#13833; 1:1000), anti-cleaved caspase-1 (#4199; 1:1000), anti-eNOS (#32027; 1:1000), anti-H3K9ac (#9649; 1:1000), anti-H3K56ac (#4243; 1:1000), anti-Histone H3 (#4499; 1:2000) and anti-β-actin (#3700; 1:1000) antibodies were from Cell signaling technologies (CST, Beverly, MA, USA). Anti-SIRT6 (ab191385; 1:2000), anti-H3 (ab1791; 1:2000) and anti-VEGF (ab46154; 1:1000) antibodies were from Abcam (Cambridge, UK).
9
Piperine Modulates PI3K/AKT and NLRP3 Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Piperine (PIP) specified to be more than 97% pure was purchased from Shanghai Winherb Medical Science Co. (Shanghai, China).13 Lactate dehydrogenase (LDH) and creatine kinase (CK) were detected by commercially available ELISA kits (Jiancheng Bioengineering Institute). The following primary antibodies were from Abcam: anti‐p‐PI3K (1:600 dilution, ab182651), anti‐t‐PI3K (1:400, ab191606), anti‐NLRP3 (1:1000, ab263899), anti‐IL‐18 (1:500, ab191860) and anti‐GAPDH (1:1000, ab37168). The following primary antibodies were obtained from Cell Signaling Technology: anti‐p‐AKT (1:800, #9145), anti‐t‐AKT (1:600, #9139), anti‐cleaved caspase‐1(1:600, #89332), anti‐pro‐caspase‐1 (1:1000, #24232), anti‐cleaved IL‐1β (1:500, #63124) and anti‐pro‐IL‐1β (1:500, #12703). Antibody against RP105 (1:600, PAB18126) was obtained from ABNOVA. The BCA protein assay kit was purchased from Pierce. Evans blue and TTC dying were purchased from Beyotime Institute of Biotechnology. LY294002 (a PI3K/AKT inhibitor) were from Sigma‐Aldrich. The miR‐383 mimic and its scrambled oligonucleotides (miR‐NC) were produced by GenePharma Co., Ltd.
10
Western Blotting Analysis of Inflammasome Components
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HK‐2 cells and renal tubule tissue were lysed in RIPA buffer (Solarbio, Beijing, China) with protease inhibitor using the following primary antibodies: anti‐TXNIP (14715, Cell Signaling Technology), anti‐NLRP3 (15101, Cell Signaling Technology), anti‐caspase‐1 (24232/3866, Cell Signaling Technology), anti‐cleaved IL‐1β (83186/63124, Cell Signaling Technology), anti‐ASC (sc‐514414, Santa Cruz), anti‐GSDMD (NBP2‐33422, Novus Biologicals, CO/ab219800, Abcam), anti‐cleaved caspase‐1 (4199/89332, Cell Signaling Technology), anti‐RBMX (14794, Cell Signaling Technology), anti‐β‐actin (sc‐81,178, Santa Cruz), and anti‐Histone3 (17168‐1‐AP, Proteintech, Wuhan, China). Western blotting was performed after incubation with a horseradish peroxidase‐conjugated anti‐rabbit/anti‐mouse secondary antibody (D110011/D110087, Sangon Biotech, Shanghai, China).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!