Cardiac enzymes, including Creatine Kinase Myocardial Bound (CK-MB, cat no. 8.05.13.0.0100, Atlas Medical, Cambridge, UK), Creatine Phosphokinase and Lactate Dehydrogenase (CPK, LDH, cat no. ab155901, ab102526 respectively, Abcam Inc., Waltham, MA, USA), and Aspartate Aminotransferase (AST, cat no. MBS2540582, MyBioSource, San Diego, CA, USA) were measured as mentioned in the manufacturer’s procedure.
Lactate dehydrogenase
Manufactured by Abcam
Sourced in United Kingdom
Lactate dehydrogenase is an enzyme that catalyzes the conversion of lactate to pyruvate and vice versa. It is involved in the glycolytic pathway and plays a role in energy production within cells.
Automatically generated - may contain errors
Lab products found in correlation
Tris base, by Merck Group (1 mentions)
Salidroside, by Merck Group (1 mentions)
Dithiotheritol, by Merck Group (1 mentions)
Glutathione reductase, by Abcam (1 mentions)
Sep pak c18 cc vac cartridge, by Waters Corporation (1 mentions)
Pencillin streptomycin, by Merck Group (1 mentions)
Rpmi 1640 media, by Merck Group (1 mentions)
Milli q water, by Merck Group (1 mentions)
Dulbecco s phosphate buffer saline, by Merck Group (1 mentions)
Radioimmunoprecipitation assay ripa buffer, by Merck Group (1 mentions)
Mr frosty freezing container, by Thermo Fisher Scientific (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Isopropanol, by Merck Group (1 mentions)
Urea, by Merck Group (1 mentions)
Iodoacetamine, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Sequence grade modified trypsin, by Promega (1 mentions)
5 protocols using lactate dehydrogenase
1
Cardiac Enzyme Measurement Protocol
2
Oxidative Stress Profiling of HL-60 Cells
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HL-60 cells (HL-60(TB) (RRID:CVCL_A794 )), RPMI-1640 media, fetal bovine serum (FBS), pencillin–streptomycin, nigerose, salidroside, sterilized filtered Dulbecco's phosphate buffer saline, trypan blue solution cell culture, DMSO, isopropanol, Tris base, urea, Hydrochloric acid (HCL), ammonium biocarbonate, acetonitrile, dithiotheritol (DTT), iodoacetamine, formic acid, radio immunoprecipitation assay (RIPA) buffer, protease inhibitor cocktail, and milli-Q water were all purchased from Sigma-Aldrich (Poole, UK). A Mr. Frosty freezing container was purchased from ThermoFisher Scientific (Waltham, MA). Certified Sep-Pak C18 cc vac cartridge was purchased from (Waters, UK). Sequence grade modified trypsin was purchased from Promega (Southampton, UK). Glutathione reductase, lactate dehydrogenase, and lipid peroxidation (MDA) assay kits were purchased from Abcam (Cambridge, UK). A protein carbonyl colorimetric assay kit was purchased from Cayman Chemical Company (Ann Arbor, MI).
3
Protein Expression Analysis of Osteoclastogenesis
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Whole cell proteins were extracted via RIPA lysis buffer (P0013B, Beyotime Co., Shanghai, China), quantified by BCA assay (Sigma-Aldrich Co), separated on NuPAGE 10%–12% polyacrylamide gels, transferred to PVDF membranes (Millipore, Billercia, MA) and blocked in 5% BSA in TBST. Incubation was performed with antibodies against Cathepsin K (Abcam, Cambridge, England, 1:1000), TRAP (Abcam:1000), MMP-9 (Cell signalling, 1:1000), Calcitonin receptor (Abcam, 1:800), SerpinB2 (Abcam, 1:800), Peroxiredoxin 6 (Abcam, 1:1000), SOD2 (Abcam, 1:2000), D4 GDI (Abcam, 1:500), Lactate Dehydrogenase (Abcam, 1:3000), OPG (Santa Cruz Biotechnology, 1:1000), RANKL (Santa Cruz Biotechnology, 1:1000), β-actin (Cell-signaling, 1:2000). After washing with TBST, blots were incubated with goat anti-rabbit, goat anti-mouse or rabbit anti-goat horseradish peroxidase-conjugated secondary antibodies previous to visualization with enhanced chemi-luminescence ECL kits (Sigma). Relative band intensities in scanned images were analyzed with Image J software.
4
Enzymatic Activity Profiling in S. aureus
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Commercially available colorimetric assay kits of enolase (Abcam), pyruvate dehydrogenase (Abcam), phosphofructokinase (Abcam), and lactate dehydrogenase (Abcam) were used to measure corresponding enzymatic activities. In brief, S. aureus cell cultures with or without treatment of AgNO3 were harvested, washed, suspended, and sonicated in lysis buffers. The protein concentration of supernatant after centrifugation was measured by BCA assay. Enzyme activities were measured according to the standard procedures provided by the manufacture. The enzymatic activity was normalized to protein concentration. The activity of alkyl hydroperoxide reductase70 , Pgl71 (link), and 6PGDH72 (link) were performed according to previous reports.
5
Subcellular Fractionation and Protein Analysis
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Cells were collected in ice-cold PBS, pelleted at 1500 rpm for 5 minutes at 4 °C, and resuspended in an ice-cold hypotonic buffer (10 mM HEPES, pH 7.2, 10 mM KCl, 1.5 mM MgCl2, 0.1 mM EGTA, 20 mM NaF, 100 mM Na3VO4) supplemented with protease inhibitor cocktail. Samples were lysed in a Dounce tissue homogenizer with glass pestle until only nuclei were visible by light microscopy. Samples were centrifuged briefly to separate the supernatant from the nuclei, and the supernatant was centrifuged at 10,000 rpm for 10 minutes and collected as the "cytoplasmic" fraction. The nuclear pellet was lysed in CST lysis buffer, sonicated using a Bioruptor bath sonicator (Diagenode), centrifuged at 10,000 rpm for 10 minutes at 4 °C, and this lysate collected as the "nuclear" fraction. Samples were resolved by SDS-PAGE, and blotted for lamin A/C (CST) or lactate dehydrogenase (Abcam) as markers for the nuclear and cytoplasmic fractions respectively.
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