Solarbio triglicerid (Tg) Komplet za ispitivanje sadržaja

Triglicerid(Tg) Komplet za ispitivanje sadržaja

Bilješka: Uzmite dva ili tri različita uzorka za predviđanje prije testa.

Operativna oprema: Microplate reader/Spectrophotometer

Catalog Number: BC0625

Veličina:100T/96S

Komponente:

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

Reagens II: 3 mL×1 bottle. Skladištenje na 4 ℃.

Reagens iii: 10 mL×1 bottle. Skladištenje na 4 ℃.

Reagens IV: 3 mL×1 bottle. Skladištenje na 4 ℃, protected from light. Reagens v: 10 mL×1 bottle. Skladištenje na 4 ℃, protected from light. Reagent VI: 10 mL×1 bottle. Skladištenje na 4 ℃, protected from light.

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

Opis proizvoda:

Triglicerid(Tg) 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.

Potrebni reagensi i oprema, ali nisu pruženi:

Spectrophotometer/microplate reader, micro glass cuvette/96 well flat-bottom plate, n-heptane, isopropyl alcohol, vodena kupelji, Podesiva pipeta, distilled water and 125 mL of empty bottle.

Postupak:

Ja. Priprema uzorka:

• Tkivo:

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 of tissue and add 1 mL of Reagent I). Centrifuga na 8000 g for 10 minutes at 4℃, supernatant is used for test.

• Bacteria:

Collect 5 million cells or bacteria into the centrifuge tube, then discard the supernatant, final add 1 mL of Reagent I. Splitting bacteria or cell with ultrasonic for 1 minuta (vlast 20%, work time 2s, interval 1s). Centrifuga na 8000 g for 10 minutes at 4℃, supernatant is used for test.

• Serum: Detect

Ii. Postupak:

1. Preheat Spectrophotometer for 30 minuta, Podesite valnu duljinu na 420 NM, Postavite nulu s destiliranom vodom.
2. Preheat water bath to 65℃.

Ime reagensa (μL)	Prazna cijev (Ab)	Standard( KAO)	Epruveta(NA)
Destilirana voda	40	–	–
Standard solution	–	40	–
TG test solution	–	–	40
Reagens ⅰ	125	125	125
Reagens ⅱ	25	25	25
Mix thoroughly after adding Reagent I, add Reagent II, shake strongly for 30 s, stand several minutes. After layering, 15 μL of the upper layer solution is taken and put it into a new EP tube.

3. Detect TG content:

Ime reagensa (μL)	Prazna cijev (Ab)	Standard (KAO)	Epruveta (NA)
Upper layer solution	15	15	15
Reagens ⅲ (ul)	50	50	50
Reagens ⅳ (ul)	15	15	15
Temeljito promiješajte, water bath at 65℃ for 3 minuta.
Reagent Ⅴ (ul)	50	50	50
Reagent Ⅵ (ul)	50	50	50
Temeljito promiješajte, water bath at 65℃ for 3 minuta.

After cooling, transfer liquid from the EP tube to a micro glass cuvette/96 well flat-bottom plate, and determine the absorbance at 420 NM.

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

Iii. Izračunavanje:

1) Serum:

Tg(MG/DL)= C×(NA- Ab)÷(KAO- Ab)×100 =(NA- Ab)÷(KAO- Ab)×100

2) Tkivo:

Koncentracija proteina:

Tg(mg/mg prot)= C×V×(NA- Ab)÷(KAO- Ab) ÷(Cpr×V)= =(NA- Ab)÷(KAO- Ab) ÷Cpr

Težina uzorka:

Tg(mg/g) = C×V×(NA- Ab)÷(KAO- Ab) ÷W=(NA- Ab)÷(KAO- Ab) ÷W

3 ) Bacteria or stanice:

Tg(U/104 ćelija) = C×(NA- Ab)÷(KAO- Ab) ÷D=(NA- Ab)÷(KAO- Ab) ÷D 1dL =100 mL;

V: The volume of reagent1, 1 ml;

C: Standard concentration, 1 mg/ mL;

CPR: Koncentracija proteina uzorka (mg/ml); W: Težina uzorka(g);

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

Bilješka:

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.

Reference

• Fletcher M J. A colorimetric method for estimating serum triglycerides[J]. Acta kemijska klinika, 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