*Assay 1 and 2 used an LDH concentration of 250 ug/ml *Assay 3, 4, and 5 and use
ID: 510998 • Letter: #
Question
*Assay 1 and 2 used an LDH concentration of 250 ug/ml
*Assay 3, 4, and 5 and used an LDH concentration of 25ug/ml
Present a table showing initial velocity data (DA340nm/min, DNADH[nM]/min, and NADH[nmol]/min) as a function of LDH concentration.
LDH (ug)/assay
6.25
2.5
1.25
0.625
0.25
A340/min
0.504
0.272
0.174
0.115
0.089
NADH[nM]/min
NADH[nmol]/min
How do i get NADH[nM]/min and NADH[nmol]/min??
PLEASE HELP
Assay 1 2 3 4 5 LDH (ug/assay) 6.25 2.5 1.25 0.625 0.25 1.5 M Tris pH 8.8 915ul 930ul 890ul 915ul 930ul 300 mM Lactate 30ul 30ul 30ul 30ul 30ul 38 mM NAD 30ul 30ul 30ul 30ul 30ul LDH 25ul 10ul 50ul 25ul 10ulExplanation / Answer
This is a Project Work based on some topic related with Lactic Acid Dehydrogenase. So, it is very important to understand few points. Hopefully, this is your topic, Lactate Dehydrogenase-Catalyzed Regeneration of NAD from NADH for Use in Enzyme-Catalyzed Synthesis.
What is Assay? An assay is an analytic procedure in laboratory for qualitatively assessing or quantitatively measuring the presence, amount, or functional activity of a target entity (the analyte). The analyte can be a drug, a biochemical substance, or a cell in an organism or organic sample. The measured entity is generally called the analyte, the measurand or the target of the assay.
LDH (ug/assay) Lactate dehydrogenase (also called lactic acid dehydrogenase, or LDH) is an enzyme found in almost all body tissues.This enzyme help us in the process of turning sugar into energy for our cells into our body for use.
NADH & NAD+ short for nicotinamide adenine dinucleotide, is an important pyridine nucleotide that functions as an oxidative cofactor in eukaryotic cells. NADH plays a key role in the production of energy through redox reactions. NAD serves as a cofactor (A substance, for example, a coenzyme or metal ion, that acts with and is essential to the activity of an enzyme) for dehydrogenases, reductases and hydroxylases, making it a major carrier of H+ and e- in major metabolic pathways such as glycolysis, the triacarboxylic acid cycle, fatty acid synthesis and sterold synthesis.
1.5 M Tris pH 8.8 is buffer being used as medium in which production of NDA is going out.
L-lactate dehydrogenase catalyzes the reduction of pyruvate to form L-lactate in the presence of NADH. It also catalyzes the oxidation of L-lactate to form pyruvate in the presence of NAD+ as an electron acceptor.
Now, main part as for your calculation,
First, you need to convert LDH in ug/ assay into nM and it can be done by dividing LDH in ug/ assay with Molecular weight of LDH (LDH can be extracted from different sources, so look into sample for source of LDH and dependingly find out the Molecular weight of LDH. Let it be LDH1(nM/ assay).
Now for calculating, NADH [nM/min], you can use two method
first is , by plotting a graph of LDH1(nM/ assay) in y axis and A340/min in x-axis, and then calculating slope at various points of A340/min as needed for your project.
second method is by, multiplying LDH1(nM/ assay) with A340/min directly.
NADH [nM/min] & NADH [nmol/min] are same.
regarding initial velocity, it can be done by various methods but sufficeint system data (apparatus and machine details like area of cross section, density of NADH etc) in which work is being carried out is not given.
Related Questions
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.