H2S Content determination reagent kit instruction manual

Micro method 100T/96S

Before formal testing, it is necessary to take 2-3 Make predictions based on samples with significant expected differencesMeasurement significance:

H2S It is a new type of gaseous signaling molecule that exists in the brain as a neurotransmitter with physiological concentrationsH2S Long term potentiation of the hippocampus in the nervous systemStrong function plays an important regulatory role and has an impact on spontaneous hypertensionoutThe process of diseases such as hemorrhagic shock and cirrhosis plays an important pathological roleThe rational effect.

Measurement principle:

H2S Compared to zinc acetateN,N-Dimethyl p-phenylenediamine reacts with ammonium iron sulfate to form methylene blue, which is present in 665nm There is maximum suction at this locationPeak absorption can be calculated by measuring its absorbance valueH2SContent.

H2S content determination kit signal series - readily available in stock

composition

Self provided instruments and supplies:

Balance, low-temperature centrifuge, enzyme-linked immunosorbent assay reader96 Orifice plate, distilled water.

The specific product parameters are subject to the parameters in the product manual received

sampleplaceReason:

1. Organization: According to organizational quality（g）Volume of extraction solution(ml)for 1：5~10The proportion of（Suggest weighing the agreement 0.1gOrganize, join 1mlextractLiquid)Perform ice bath homogenization, and then 10000g，4℃centrifugation 10minTake the supernatant and place it on ice for testing.

2. Bacteria and fungi: according to the number of cells（10⁴ Individual: volume of extraction solution（ml）for 500~1000：1 The proportion of（suggestion 500 Ten Thousand Cells Plusenter 1ml extract solution）Ice bath ultrasonic disruption of cells（power 300w, Ultrasound 3 Seconds, intervals 7 Second, total time 3min）And then 10000g， 4℃, centrifugation 10minTake the supernatant and place it on ice for testing.

3. Serum (plasma)Direct measurement.

Operation table:

1Preheating of ELISA reader 30minAdjust the wavelength to 665nm.

2、operation sheet

H2S Content calculation formula:

use 96 The calculation formula for orifice plate measurement is as follows

The standard curve regression equation is:y =0.0022x，R2 =0.9988

（1）Organize samples

a.Calculated based on protein concentration

H2S（nmol/mg prot）=b.Calculated based on sample weight H2S（nmol/g Fresh weight)=

(3)cell

H2S（nmol/10⁴ cell）=

ΔA ×V Anti General÷(V sample×Cpr)=681.8×ΔA÷Cpr

0.0022

ΔA ×V Anti General÷(V sample÷V Sample General×W)= 681.8×ΔA÷W

0.0022

ΔA ×V Anti General÷（V sample÷V Sample General×cell count(ten thousand)）=681.8×ΔA÷Number of cells (10000)）

0.0022

（2）liquid sampleH2S（nmol/ml）=

ΔA ×V Anti General÷V sample=681.8×ΔA

0.0022

V Counter total: total volume of reaction,0.225ml；V Sample: volume of sample in the reaction,0.15ml；V Sample total: Add the volume of extraction solution,1ml；W：Sample quality,g；CprProtein concentration,mg/ml

Precautions:Zui low detectionoutLimited to 1nmol/ml.

H2S content determination kit signal series - readily available in stock