Solarbio Triglycerid (Tg) Inhaltstest-Kit

Triglycerid(Tg) Inhaltstest-Kit

Notiz: Nehmen Sie vor dem Test zwei oder drei verschiedene Proben zur Vorhersage.

Betriebsausrüstung: Microplate reader/Spectrophotometer

Katalognummer: BC0625

Größe:100T/96S

Komponenten:

Reagenz I: Selbstbereitete Reagenz, hinzufügen 80 mL of n-heptane and 80 mL of isopropyl alcohol to an empty glass bottle. Seal and mix well, storage at 4℃.

Reagenz II: 3 mL×1 bottle. Lagerung bei 4℃.

Reagenz III: 10 mL×1 bottle. Lagerung bei 4℃.

Reagenz IV: 3 mL×1 bottle. Lagerung bei 4℃, vor Licht geschützt. Reagenz V: 10 mL×1 bottle. Lagerung bei 4℃, vor Licht geschützt. Reagenz VI: 10 mL×1 bottle. Lagerung bei 4℃, vor Licht geschützt.

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

Produktbeschreibung:

Triglycerid(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.

Erforderliche, aber nicht bereitgestellte Reagenzien und Ausrüstung:

Spektrophotometer/Mikroplatten-Reader, Mikroglasküvette/96-Well-Platte mit flachem Boden, n-heptane, isopropyl alcohol, Wasserbad, verstellbare Pipette, distilled water and 125 mL of empty bottle.

Verfahren:

ICH. Probenvorbereitung:

• Gewebe:

Die Eisbad-Homogenität wird gemäß dem Verhältnis der Gewebemasse durchgeführt (G): Reagent Ⅰ volume (ml) = 1: 5~ 10 (es wird vorgeschlagen, herum zu nehmen 0.1 G Gewebe und hinzufügen 1 ml Reagenz I). Zentrifuge bei 8000 G für 10 Minuten bei 4 ℃, supernatant is used for test.

• Bakterien:

Sammeln 5 million cells or bacteria into the centrifuge tube, then discard the supernatant, final add 1 ml Reagenz I. Splitting bacteria or cell with ultrasonic for 1 Minute (Leistung 20%, work time 2s, interval 1s). Zentrifuge bei 8000 G für 10 Minuten bei 4 ℃, supernatant is used for test.

• Serum: Erkennen

II. Verfahren:

1. Preheat Spectrophotometer for 30 Protokoll, Passen Sie die Wellenlänge an 420 nm, Mit destilliertem Wasser auf Null stellen.
2. Preheat water bath to 65℃.

3. Detect TG content:

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

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

III. Berechnung:

1) Serum:

Tg(mg/dL)= C×(BEI- AB)÷(ALS- AB)×100 =(BEI- AB)÷(ALS- AB)×100

2) Gewebe:

Proteinkonzentration:

Tg(mg/mg prot)= C×V×(BEI- AB)÷(ALS- AB) ÷(Cpr×V)=(BEI- AB)÷(ALS- AB) ÷ CPR

Probengewicht:

Tg(mg/g) = C×V×(BEI- AB)÷(ALS- AB) ÷W=(BEI- AB)÷(ALS- AB) ÷ w

3 ) Bacteria or Zellen:

Tg(U/104 -Zelle) = C×(BEI- AB)÷(ALS- AB) ÷D=(BEI- AB)÷(ALS- AB) ÷D 1dL =100 mL;

V: The volume of reagent1, 1 ml;

C: Standard concentration, 1 mg/ mL;

Cpr: Proteinkonzentration der Probe (mg/ml); W: Probengewicht(G);

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

Notiz:

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.

Verweise

• Fletcher M J. A colorimetric method for estimating serum triglycerides[J]. Acta Chemical Clinic, 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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Technische Spezifikationen:

The detection limit: 0.0372 mg/ml

Linear range: 0.0625-3 mg/ml