293FT cells were co-transfected with survivin luciferin reporter plasmid (1.2 μg) and CMV-LacZ reporter plasmid (0.3 μg) using Lipofectamine 2000 transfection reagent (Invitrogen, Canada) according to the manufacturer's instructions. Total protein was extracted at 24 hours post transfection and subjected to luciferase and β-galactosidase detection using a luciferase assay kit (Promega, Canada) and a β-galactosidase enzyme assay kit (Promega, Canada) respectively, according to the manufacturer's instructions.
β galactosidase enzyme assay kit
Manufactured by Promega
Sourced in United States
The β-galactosidase enzyme assay kit is a laboratory tool used to measure the activity of the β-galactosidase enzyme. The kit provides the necessary reagents and protocols to quantify the level of β-galactosidase in a sample. The core function of this product is to enable the analysis and quantification of β-galactosidase in various experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Luciferase assay kit, by Promega (1 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Sirtinol, by Merck Group (1 mentions)
Rabbit anti sirt1 polyclonal antibody, by Santa Cruz Biotechnology (1 mentions)
Sa β galactosidase staining kit, by Merck Group (1 mentions)
Horseradish peroxidase hrp conjugated igg, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti β actin polyclonal antibody, by Merck Group (1 mentions)
Ang 2, by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Resveratrol, by Merck Group (1 mentions)
Luciferase assay system, by Promega (1 mentions)
Biotek synergy ht, by Agilent Technologies (1 mentions)
6 protocols using β galactosidase enzyme assay kit
1
Transfection and Reporter Assay
2
Senescence and Bone Matrix Analysis
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SAβG activity assay was performed by β‐galactosidase enzyme assay kit (Promega, San Luis Obispo, CA, USA); the absorbance at 420 nm was measured according to the manufacturer's instructions. Bone tissue β‐galactosidase staining was also performed according to a method published previously.(29 , 30 ) Senescent cells were identified as blue‐stained cells by standard light microscopy. After SAβG staining, slides were further stained with Col‐1a and osteocalcin (antiosteocalcin, Ms, monoclonal, #ab13418; Abcam) using standard immunostaining.
3
Beta-Galactosidase Enzyme Assay for SABG Activity
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The SABG activity assay was performed using a β‐galactosidase enzyme assay kit (Promega, San Luis Obispo, CA, USA), measuring the absorbance at 420 nm according to the manufacturer's instructions. Isolation of proteins from cells and bone tissues for SABG activity measurements were described previously.20 , 22
4
Quantifying SAβG Enzymatic Activity
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SAβG activity assay was performed by β-galactosidase enzyme assay kit (Promega) measured the absorbance at 420 nm according to manufacturer’s instruction as we described previously(19,20).
5
Resveratrol Modulation of SIRT1 in Cell Senescence
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Ang II, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), polyclonal rabbit anti-β-actin antibody, resveratrol, SA β-galactosidase staining kit, sirtinol, and whey protein (W1500) were obtained from Sigma-Aldrich Co. (USA). The β-galactosidase enzyme assay kit and luciferase assay system were obtained from Promega (USA). Rabbit anti-SIRT1 polyclonal antibody and horseradish peroxidase (HRP)-conjugated IgG were obtained from Santa Cruz Biotechnology (USA).
6
Measuring LacZ reporter activity in E. coli and P. aeruginosa
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LacZ reporter assay was performed in both E. coli and in P. aeruginosa strains as previously described (Cugini et al., 2007 (link)). Briefly, overnight culture of E. coli containing the pEAL08-2 reporter construct was diluted to OD600nm of 0.05, grown in the presence of quinolones for 2 h, and harvested by centrifugation. P. aeruginosa strains with pEAL08-2 were grown overnight before harvest. Cells were lysed and the LacZ activity was measured using a β-galactosidase enzyme assay kit (Promega, Madison, WI, USA). The LacZ product was measured at 420 nm on a plate reader (BioTek Synergy HT; BioTek, Winooski, VT, USA).
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