- Catalogue No.
K127
- Size
48T(32S)/ 96T(80S)
- Detection range
1.77-40 μmol/L
- Intended use
The kit can be used to detect the concentration of total bile acid (TBA) in serum, plasma and animal tissue samples.
- Kit component
|
Item
|
Size（48T）
|
Size（96T）
|
|
Chromogenic Agent A
|
12 mL
|
24 mL
|
|
Chromogenic Agent B
|
3 mL
|
6 mL
|
|
0.2 mmol/L Standard
|
2 mL
|
2 mL
|
- Storage
The kit should be stored at 2-8℃ shading light for 3 months.
- Principle of the Assay
With S-NAD+ as hydrogen receptor, 3α-hydroxy steroid dehydrogenase catalyzed the dehydrogenation of bile acids to produce 3-ketone steroids, transforming S-NAD+ into S-NADH. Meanwhile, NADH was used as hydrogen donor. 3α-hydroxy steroid dehydrogenase catalyzed the production of bile acids from 3-ketone steroids. Through the enzyme cycle reaction, S-NADH is continuously generated, which has the maximum absorption peak at 405 nm. Measure the OD value at 405 nm and the changes of absorbance is proportional to the concentration of bile acid.
- Additional Materials Required
Instruments:
Microplate reader (400-410 nm, optimum wavelength: 405 nm), Micropipettor, Centrifuge, Incubator, Vortex mixer
Reagents:
Double distilled water, Normal saline (0.9% NaCl)
- Calculation
The standard curve:
1. Average the duplicate reading for each standard.
2. Subtract the mean ΔABlank/min (ΔA/min value when the standard concentration is 0) of the blank (Standard #1) from all standard readings. This is the ΔA405 (ΔAStandard/min– ΔABlank/min).
3. Plot the standard curve by using ΔA405 of standard and correspondent concentration as y-axis and x-axis respectively. Create the standard curve (y = ax+ b) with graph software (or EXCEL).
The sample:
1.Serum (plasma) sample: TBA content(μmol/L) = (ΔA405- b) ÷ a× f
2.Animal tissue sample: TBA content(μmol/gprot) = (ΔA405- b) ÷ a× f ÷ Cpr
[Note]
ΔA405: △ASample/min-△ABlank/min
f: Dilution factor of the sample before tested.
Cpr：The concentration of protein in sample, gprot/L.
- Notes
1.This assay kit is for Research Use Only. We will not response for any arising problems or legal responsibilities causing by using the kit for clinical diagnosis or other purpose.
2.Please read the instructions carefully and adjust the instruments before the experiments. Please follow the instructions strictly during the experiments.
3.Protection methods must be taken by wearing a lab coat and latex gloves during the experiment.
4.If the concentration of substance is not within the detection range exactly, an extra dilution or concentration should be taken for the sample.
5.It is recommended to take a pre-test if your sample is not listed in the instruction book.
6.The experimental results are closely related to the situation of reagents, operations, environment and so on. Elabscience will guarantee the quality of the kits only, and NOT be responsible for the sample consumption caused by using the assay kits. It is better to calculate the possible usage of sample and reserve sufficient samples before use.
