Total cholesterol (TC) content detection kit

Product Introduction:

Total cholesterol（Total Cholesterol，TC）It refers to the total amount of cholesterol contained in all lipoproteins, including free cholesterol and cholesterol esters.

Using esterase catalyzed hydrolysis of cholesterol esters to generate free cholesterol（FC）And free fatty acids（FFA）Thereby converting cholesterol estersfor FCFurther utilize cholesterol oxidase catalysis FCOxidation, generation△4-Cholesterenone and H₂O₂Finally, utilize peroxidase catalysisH₂O₂ oxidation 4-Aminoantipyrine and phenol form red quinone compounds, which are present in 500nmThere is a characteristic absorption peak, and its color depth is related toTCThe content is directly proportional.

Instruments and reagents required for the experiment：

Visible spectrophotometer/ELISA reader, balance, low-temperature desktop centrifuge, micro glass colorimetric dish/96Orifice plate, adjustable pipette

EPTube, distilled water, isopropanol.

Product Content:

Extraction solution: Isopropanol 100mL×1Bottle, bring your own,4℃保存；working fluid: Liquid 30mL×1Bottle,4℃保存；

Standard product: Powder×1Support,10mg cholesterol,4Store at ℃. Add before use517MLThe extraction solution is dissolved by shaking, which is50Mmol/mLThe cholesterol standard solution.

Operation steps:

1、 Sample processing:

1. Organization: According to organizational quality（g）Volume of extraction solution(mL)for 1：5~10 The proportion of（Suggest weighing the agreement 0.1g Organize, join 1mL

Extract the liquid and homogenize it in an ice bath.10000g，4℃ centrifugation10minTake the supernatant and place it on ice for testing.

2. Bacteria and fungi: according to the number of cells（104 a）Volume of extraction solution（mL）for 500~1000：1 The proportion of（suggestion 500 ten thousandCell addition 1mL extract solution）Ice bath ultrasonic disruption of cells（power 300w, Ultrasound 2 Seconds, intervals 3 Second, total time 3min）；then 10000g，4℃ centrifugation10minTake the supernatant and place it on ice for testing.

3. Serum (plasma)Sample: Direct measurement.2、 Measurement steps:

1、 spectrophotometer/Preheating of ELISA reader 30minAbove, adjust the wavelength to 500nmAdjust distilled water to zero.

2、 Translate into English 50μmol/mLDilute the standard solution with the extraction solution to 2.5、1.25、0.625、0.3125、0.15625The0.078μmol/mLThe standard solution is ready for use.

3、 Operation table:(In centrifuge tubes or 96 Add the following reagents to the orifice plate in sequence)

Total cholesterol content calculation

1、 Drawing of standard curve:

Based on the concentration of each standard solution xAxis, its corresponding△AThe standard is yAxis, draw standard curve, obtain standard equation y=kx+b， Translate into English△AEnter the equation to obtain x（μmol/mL）

2、 Calculation of total cholesterol content:

1. Serum (plasma)middle FC Content calculation: TC Content（μmol/dL）=x×100.

2. Within the organization FC Content calculation:

1. Calculated based on sample protein concentration

TCContent（μmol/mg prot）=x×Vextract÷（Cpr×VExtract)=x÷Cpr.

2. Calculated based on the fresh weight of the sample

TCContent（μmol/gFresh weight)=x×Vextract÷W= x÷W

3. In cells and bacteria TC Content calculation:

TCcontent(μmol/104 cell)=x×Vextract÷500=0.002x.

100：1dL=100mL；VExtraction: Add the volume of extraction solution to the sample,1mL；W: Sample quality,g；500:500Number of 10000 cells.

Precautions

When△Agreater than 1.5 It is recommended to dilute the sample with extraction solution before conducting the measurement.

Shanghai Yuchun Biotechnology Co., Ltd. is a technology enterprise jointly founded by personnel with backgrounds in life sciences and biotechnology. The company focuses on primary cells, tumor cells, and cell culture related reagent products, providing technical support and services for universities, food, medicine, biological research, and other fields.

Since its establishment, the company has provided customized cell products and complete cell culture solutions to customers through continuous technological innovation and product research and development. Through active exploration, research and development, deepening and enriching of products, there are currently more than 200 types of basic culture medium products, more than 300 cell lines, and nearly 600 types of primary cells. We always take "developing science and technology, caring for public life and health" as our driving force. Through technological innovation, we extensively attract high-tech talents, continuously improve our product system and enrich our product chain, actively expand our market and application areas, and establish a competitive "Yuchun" brand in the market.

Precautions

1. For your safety and health, please wear lab coats and disposable gloves and masks when operating;

2. To maintain the excellent performance of this product, please be sure to store it according to the recommended storage conditions;

3. This product is only for scientific research or further production use and cannot be used for clinical diagnosis or treatment.

Total cholesterol (TC) content detection kit