Drying studies were performed to evaluate the presence of residual ethanol in the final eluate. TruTips were subjected to the following mixtures containing ethanol: 95% ethanol/Lysis Buffer F (Akonni, Cat# 301-20006), Akonni Wash Buffer J, and Wash Buffer K (Akonni, Cat# 301-20027). After binding and wash cycles, the TruTips were dried for either 0, 15, 30, 45, or 60 seconds using the pressurized air pump on the workstation (n=3 per condition). Alternatively, the TruTips were dried by either 30 or 60 aspiration and dispense cycles in an empty well (n=3 per condition). Elution Buffer A2 was then passed through the matrix and collected. The eluate was analyzed with the NanoDrop 1000 (ThermoFisher, Waltham, MA) for ethanol concentration using a colorimetric assay (NAD-ADH Reagent, Sigma, St. Louis, MO, Cat# N7160-10X16ML). Samples with varied drying conditions were compared to standards prepared with known quantities of ethanol in Elution Buffer A2 (50% – 0.01%) to determine the ethanol content in the final eluate.
Nad adh reagent
Manufactured by Merck Group
Sourced in United States
NAD-ADH Reagent is a laboratory product manufactured by Merck Group. It is a reagent used for the detection and quantification of nicotinamide adenine dinucleotide (NAD) and alcohol dehydrogenase (ADH) in various biological samples. The product provides the necessary components for the enzymatic analysis of these analytes.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using nad adh reagent
1
Evaluating Ethanol Residuals in TruTips
2
Fly Ethanol Absorption and Metabolism
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Flies were frozen in liquid nitrogen immediately following 10min of 50% ethanol exposure for absorption studies and 50min post-exposure for metabolism studies. Flies were homogenized in 0.5M glycine (pH 9, 200µl for 6 flies). 2-fold dilution was performed on 100µl of supernatant following centrifugation at 12,000rpm for 5min. Following a second round of centrifugation (12,000rpm for 5min) ethanol concentrations were measured in fly homogenate supernatant using NAD-ADH Reagent (N7160, Sigma-Aldrich, St. Louis, MO, USA). The reaction was placed in a Synergy HTX plate reader (BioTek, Winooski, VT,USA) and measured every 5 minutes for 30minutes at 340nm. To calculate the ethanol concentration in flies one male fly was estimated to be 0.75mg based on average mass recorded during experiments. Calculations involved extrapolating XmM ethanol content at 15min and a final calculation of
) =mM/fly , where DF is dilution factor and Z is total mass of flies within each sample.
) =mM/fly , where DF is dilution factor and Z is total mass of flies within each sample.
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