DNCB was obtained from Sigma-Aldrich (St. Louis, MO). Imiquimod cream [5 weight % (wt %)] was purchased from 3M Pharmaceuticals (Aldara; Leicestershire, UK). Dex cream was purchased from CR Sanjiu (Shenzhen, China). Tacrolimus ointment was purchased from Astellas (Toyama, Japan). Penicillin-streptomycin, PBS, and fetal bovine serum (FBS) were purchased from Thermo Fisher Scientific (USA). HA was obtained from Bloomage Freda Biopharm Co. Ltd. (Shandong, China). CTP was supplied by Jellice Co. Ltd. (Miyagi, Japan). CRISPR-Cas9 protein was purchased from GenScript (Nanjing, China). T7E1 enzyme was obtained from New England Biolabs (Beijing, China). Lipofectamine CMAX Transfection Reagent was bought from Thermo Fisher Scientific (Germany). sgNLRP3 was designed by online tools (http://crispr.mit.edu/ and http://chopchop.cbu.uib.no/ ) and then prepared using the HiScribe T7 Quick High Yield RNA Synthesis Kit (New England Biolabs, MA, USA). Primers for in vitro transcription of gRNAs are listed in table S1. Primary antibodies used in this project included the following: Anti-NLRP3 (1:1000; ER1706-72) antibody was obtained from HUABIO (Hangzhou, China), anti–caspase-1 p10 (1:1000; sc-514) and anti–glyceraldehyde-3-phosphate dehydrogenase antibodies were obtained from Santa Cruz Biotechnology (CA, USA), and anti–IL-1β (1:1000; ab200478) was obtained from Abcam (Shanghai, China).
Anti glyceraldehyde 3 phosphate dehydrogenase antibody
Manufactured by Santa Cruz Biotechnology
Sourced in China, United States
Anti–glyceraldehyde-3-phosphate dehydrogenase antibodies are a type of antibody that specifically binds to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein. GAPDH is an enzyme involved in the glycolysis pathway and is commonly used as a reference or loading control in various biochemical and cell biology experiments.
Automatically generated - may contain errors
Lab products found in correlation
Anti caspase 1 p10, by Santa Cruz Biotechnology (1 mentions)
T7e1 enzyme, by New England Biolabs (1 mentions)
Anti nlrp3, by Huabio (1 mentions)
Hiscribe t7 quick high yield rna synthesis kit, by New England Biolabs (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Anti il 1β, by Abcam (1 mentions)
Imiquimod cream, by 3M (1 mentions)
Crispr cas9 protein, by GenScript (1 mentions)
Lipofectamine cmax transfection reagent, by Thermo Fisher Scientific (1 mentions)
X omat 2000a processor, by Kodak (1 mentions)
Enhanced chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Fura 2 am, by Merck Group (1 mentions)
Collagenase type 1, by Merck Group (1 mentions)
Hepes, by Merck Group (1 mentions)
4 protocols using anti glyceraldehyde 3 phosphate dehydrogenase antibody
1
CRISPR-Mediated NLRP3 Knockdown Assay
2
Western Blot Protein Detection
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Samples were electrophoresed and transferred onto nitrocellulose membranes. Membranes were incubated with anti-inducible nitric oxide synthase (iNOS) or anti-glyceraldehyde 3-phosphate dehydrogenase antibodies (Santa Cruz Biotechnology, Dallas, TX, USA) and anti-rabbit horseradish peroxidase conjugated secondary antibodies (Santa Cruz). Enhanced chemiluminescent substrate (Thermo, Mississauga, ON, Canada) was added directly onto each membrane and developed using radiographic film on a Kodak X-Omat 2000a processor (Windsor, CO, USA).
3
Biochemical Analysis of SGLT2 and Glucose Transporters
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CANA, a SGLT2i, was kindly provided by the Mitsubishi Tanabe Pharma Corporation (Osaka, Japan). All other reagents were purchased from Cell Signaling Technology, Inc. (Danvers, MA) unless otherwise indicated. Antibodies against SGLT2 were purchased from Abcam plc (ab137207 and ab85626, Kenbridge, England), Proteintech Group, Inc. (24654-1-AP, Rosemont, IL), and Cell Signaling Technology, Inc. (#14210). Antibodies against glucose transporter (GLUT) 4, GLUT5, GLUT6, anti-mitochondrial pyruvate dehydrogenase kinase, hydroxymethylglutaryl-CoA lyase, and phosphate- AMP-activated protein kinase (AMPK) α2 (Ser345) were purchased from Abcam plc. Antibodies against GLUT1, GLUT2, GLUT3, caspase8, caspase3, carnitine palmitoyltransferase (CPT) 1A, CPT2, acyl-CoA synthetase long-chain family member/fatty acid-CoA ligase, and solute carrier family 25 member 2 were purchased from Proteintech Group, Inc. Actin antibody was purchased from Sigma-Aldrich Co. LLC (St. Louis, MO). Anti-glyceraldehyde-3-phosphate dehydrogenase antibody was purchased Santa Cruz Biotechnology (Dallas, TX). Jurkat (Human) whole cell lysate was purchased from Abcam plc. Primary human hepatocytes (LHum17003) was purchased from BIOPREDIC International (Saint-Grégoire, France).
4
Isolation and Analysis of Cx43 Protein
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Type I collagenase, Joklik modified Eagle medium, HEPES, fura-2/AM, 18 and beta-glycyrrhetinic acid were purchased from Sigma Chemical Co; anti-glyceraldehyde-3-phosphate dehydrogenase antibody, polyclonal and monoclonal anti-Cx43 antibodies as well as the corresponding horseradish peroxidase-conjugated secondary antibodies were obtained from Santa Cruz Biotech. (Santa Cruz, CA). All the chemicals were dissolved in distilled water except fura-2/AM, which was dissolved in diethyl sulphoxide (DMSO).
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