Triglyceride (TG) content detection kit

Attention: Before formal testing, it is necessary to take2-3Make predictions based on samples with significant expected differences.

Reagent content:

Visible spectrophotometry

Reagent 1: Bring your own, take 100 mL Glass empty bottle, add 45mLHeptane,45mLIsopropanol, cover tightly and mix well,4℃保存。

Reagent 2: Liquid 7mL×1Bottle,4℃保存。Reagent 3: Liquid 30mL×1Bottle,4℃保存。

Reagent 4: Liquid 10mL×1Bottle,4Store in the dark at ℃.Reagent 5: Liquid 30mL×1Bottle,4Store in the dark at ℃.Reagent 6: Liquid 30mL×1Bottle,4Store in the dark at ℃.

Standard product: Powder×1Bottle, add before use 5mLReagent 1, i.e 1mg/mLTriglyceride standard solution,4℃保存。

Product Description:

TGIt is a fatty molecule formed by long-chain fatty acids and glycerol, which is not only the main component of the cell membrane, but also an important respiratory substrate. measurePrinciple of determination: Extract with isopropanol TG，KOHsaponification TGAfter hydrolysis, glycerol and fatty acids are generated, and periodic acid oxidizes glycerol to produce formaldehyde, Formaldehyde condenses with acetylacetone in the presence of chloride ions to form a yellow substance 420 nmThere is a characteristic absorption peak, and the depth of its color is related to TGThe content is directly proportional.

Self provided instruments and supplies:

Visible spectrophotometer, water bath, adjustable pipette1mLGlass colorimetric dish, double distilled water, and 100mLGlass empty bottle.

operating steps:

1TGExtraction of:

1、 Organization: According to organizational quality（g）Reagent 1 volume(mL)for1：5~10The proportion of（Suggest weighing the agreement0.1gOrganize, join1mLReagent 1) Perform ice bath homogenization,8000g，4℃ centrifugation10minTake the supernatant for testing.

2、 Cells and bacteria: collect first 400-500Transfer 10000 cells or bacteria into a centrifuge tube, discard the supernatant, and add 1mLReagent 1: Ultrasonic fragmentation

1min（intensity 20%, Ultrasound 2sStop, stop 1s），8000g， 4℃ centrifugation10minTake the supernatant for testing.

3、 Serum (slurry) and other liquid samples: direct measurement. 2、 Measurement operation:

1. Preheating of spectrophotometer 30minAdjust the wavelength to 420 nmAdjust distilled water to zero.

2. Preheat the water bath to 65℃。

After adding reagent one, mix thoroughly, then add reagent two and shake vigorously 30sLet it stand and take the upper solution after layering 75µLPlaced in a new locationof EPIn the management.

3. Triglyceride content determination:

取出 EP Tube, after cooling, in 420nmColor comparison, denoted as AEmpty,ABiao and AMeasure.Note: Blank tubes and standard tubes only need to be measured once.

3、 Calculation formula:

1. Triglyceride content in serum (plasma):

TG Content（mg/dL）=Creference standard×（ATest -AEmpty)÷（AStandard -AEmpty)×100

=100×（ATest -AEmpty)÷（AStandard -AEmpty)

C Standard substance: concentration of standard substance,1mg/mL；100Unit conversion factor,1dL=100 mL.

2. Triglyceride content in the organization:

(1)Calculated based on sample protein concentration

TG Content（mg/mg prot）=C reference standard×V×（ATest -AEmpty)÷（AStandard -AEmpty)÷（Cpr ×V）

=（ATest -AEmpty)÷（AStandard -AEmpty)÷ Cpr

(2)Calculated based on the fresh weight of the sample

TG Content（mg/gFresh weight)=Creference standard×V×（ATest -AEmpty)÷（AStandard -AEmpty)÷W

=（ATest -AEmpty)÷（AStandard -AEmpty)÷ W

CStandard product:1mg/mL；Cpr: Sample protein concentration,mg/mL；W: Fresh weight of the sample,g ;VAdd the volume of reagent one, 1mL.

3. Triglyceride content in cells and bacteria:

TG Content（mg/104 cell）=C reference standard×（ATest -AEmpty)÷（AStandard -AEmpty)÷Bacterial or cellular density

=（ATest -AEmpty)÷（AStandard -AEmpty)÷Bacterial or cellular density

C Standard substance: concentration of standard substance,1mg/mLBacterial or cellular density:104 cell/mL

Notes:

1. There are volatile substances in the reagent kit, gloves and masks should be worn during the experiment, and the reagent bottle cap should be promptly closed after opening.

2. After adding reagent 2, it is necessary to shake vigorously to fully extract triglycerides from the test solution.

3. The detection limit is 100µ mol/L.

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.

Triglyceride (TG) content detection kit