Trigliceride Solarbio (T.G) Kit di analisi del contenuto

Trigliceridi(T.G) Kit di analisi del contenuto

Nota: Prendi due o tre campioni diversi per la previsione prima del test.

Attrezzatura operativa: Microplate reader/Spectrophotometer

Numero di catalogo: BC0625

Misurare:100T/96S

Componenti:

Reagente I: Self-provided reagent, aggiungere 80 mL of n-heptane and 80 mL of isopropyl alcohol to an empty glass bottle. Seal and mix well, storage at 4℃.

Reagente II: 3 mL×1 bottle. Conservazione a 4 ℃.

Reagente III: 10 mL×1 bottle. Conservazione a 4 ℃.

Reagente IV: 3 mL×1 bottle. Conservazione a 4 ℃, protetto dalla luce. Reagente V: 10 mL×1 bottle. Conservazione a 4 ℃, protetto dalla luce. Reagente VI: 10 mL×1 bottle. Conservazione a 4 ℃, protetto dalla luce.

Standard: powder ×1 bottle, aggiungere 5 mL of Reagent I before use. 1 mg/mL triglyceride standard solution, storage at 4℃.

Descrizione del prodotto:

Trigliceridi(T.G) is a fat molecule formed by long-chain fatty acids and glycerol, which is not only the main component of cell membrane, but also an important respiratory substrate. The TG is extracted with isopropyl alcohol, then hydrolysis to glycerol and fatty acids after saponification of TG by KOH. Glycerol is oxidized by periodic acid to form formaldehyde. Condensation of formaldehyde and acetylacetone to form yellow components in presence of chloride ions. The yellow component has a characteristic absorption at 420 nm and proportional to the TG content.

Reagenti e attrezzature necessari ma non forniti:

Lettore spettrofotometro/micropiastre, microcuvetta in vetro/piastra a fondo piatto da 96 pozzetti, n-heptane, isopropyl alcohol, bagnomaria, pipetta regolabile, distilled water and 125 mL of empty bottle.

Procedura:

IO. preparazione del campione:

• Tessuto:

Ice-bath homogeneity is conducted according to the ratio of tissue mass (G): Reagent Ⅰ volume (ml) = 1: 5~10 (it is suggested to take about 0.1 g di tessuto e aggiungere 1 ml di reagente I). Centrifugare a 8000 g for 10 minuti a 4 ℃, supernatant is used for test.

• Bacteria:

Raccogliere 5 million cells or bacteria into the centrifuge tube, then discard the supernatant, final add 1 ml di reagente I. Splitting bacteria or cell with ultrasonic for 1 minuto (energia 20%, work time 2s, interval 1s). Centrifugare a 8000 g for 10 minuti a 4 ℃, supernatant is used for test.

• Siero: Detect

II. Procedura:

1. Preheat Spectrophotometer for 30 minuti, regolare la lunghezza d'onda su 420 nm, azzerare con acqua distillata.
2. Preheat water bath to 65℃.

3. Detect TG content:

Dopo il raffreddamento, transfer liquid from the EP tube to a micro glass cuvette/96 well flat-bottom plate, and determine the absorbance at 420 nm.

Nota: Blank tube and standard tube only need to be measured once.

III. Calcolo:

1) Siero:

T.G(mg/dL)= C×(A- AB)÷(COME- AB)×100 =(A- AB)÷(COME- AB)×100

2) Tessuto:

Concentrazione proteica:

T.G(mg/mg prot)= C×V×(A- AB)÷(COME- AB) ÷(Cpr×V)=(A- AB)÷(COME- AB) ÷Cpr

Peso del campione:

T.G(mg/g) = C×V×(A- AB)÷(COME- AB) ÷W=(A- AB)÷(COME- AB) ÷W

3 ) Bacteria or cellule:

T.G(Cella U/104) = C×(A- AB)÷(COME- AB) ÷D=(A- AB)÷(COME- AB) ÷D 1dL =100 mL;

V: The volume of reagent1, 1 ml;

C: Standard concentration, 1 mg/ mL;

Cpr: Concentrazione proteica del campione (mg/ml); W: Peso del campione(G);

D: Density of bacteria or cell, 104 cell/mL.

Nota:

1. There are volatile substances in the kit. Gloves and masks should be worn during the experiment. The reagent bottle cap should be closed in time after
2. After the addition of Reagent Ⅱ, it is necessary to repeatedly and violently vibrate, so that the triglyceride in the test solution can be fully extracted, and the oscillation amplitude, time, repeated times and waiting for stratification time should be
3. In order to ensure the repeatability of the test, the cooling time after each water bath should be
4. If the OD value of the test tube is greater than 1.5, it is recommended to dilute the sample with Reagent I properly before testing, and multiply it by the corresponding dilution multiple during calculation.

Riferimenti

• Fletcher M J. A colorimetric method for estimating serum triglycerides[J]. Clinica Chimica Acta, 1968, 22(3): 393-397.
• Hercules D M, Sheehan T L. Chemiluminescent determination of serum glycerol and triglycerides[J]. Analytical chemistry, 1978, 50(1):22-25.

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Technical Specifications:

The detection limit: 0.0372 mg/ml

Linear range: 0.0625-3 mg/ml