Triglyceride(TG) Content Assay Kit
Notiz: Nehmen Sie vor dem Test zwei oder drei verschiedene Proben zur Vorhersage.
Betriebsausrüstung: Microplate reader/Spectrophotometer
Catalog Number: BC0625
Größe:100T/96S
Komponenten:
Reagenz I: Self-provided reagent, 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℃.
Reagent IV: 3 mL×1 bottle. Lagerung bei 4℃, vor Licht geschützt. Reagent V: 10 mL×1 bottle. Lagerung bei 4℃, vor Licht geschützt. Reagent 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:
Triglyceride(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:
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 Reagenz I). Zentrifuge bei 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 Reagenz I. Splitting bacteria or cell with ultrasonic for 1 Minute (power 20%, work time 2s, interval 1s). Zentrifuge bei 8000 g for 10 minutes at 4℃, supernatant is used for test.
- Serum: Detect
II. Verfahren:
- Preheat Spectrophotometer for 30 Protokoll, adjust the wavelength to 420 nm, Mit destilliertem Wasser auf Null stellen.
- Preheat water bath to 65℃.
Reagent name (μL) | Blank tube (AB) | Standard tube( ALS) | Test tube(BEI) |
Distilled water | 40 | – | – |
Standardlösung | – | 40 | – |
TG test solution | – | – | 40 |
Reagent Ⅰ | 125 | 125 | 125 |
Reagent 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. |
- Detect TG content:
Reagent name (μL) | Blank tube (AB) | Standard tube (ALS) | Test tube (BEI) |
Upper layer solution | 15 | 15 | 15 |
Reagent Ⅲ (uL) | 50 | 50 | 50 |
Reagent Ⅳ (uL) | 15 | 15 | 15 |
Mix thoroughly, water bath at 65℃ for 3 Protokoll. | |||
Reagent Ⅴ (uL) | 50 | 50 | 50 |
Reagent Ⅵ (uL) | 50 | 50 | 50 |
Mix thoroughly, water bath at 65℃ for 3 Protokoll. |
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. Calculation:
1) Serum:
TG(mg/dL)= C×(BEI- AB)÷(ALS- AB)×100 =(BEI- AB)÷(ALS- AB)×100
2) Gewebe:
Protein concentration:
TG(mg/mg prot)= C×V×(BEI- AB)÷(ALS- AB) ÷(Cpr×V)=(BEI- AB)÷(ALS- AB) ÷Cpr
Sample weight:
TG(mg/g) = C×V×(BEI- AB)÷(ALS- AB) ÷W=(BEI- AB)÷(ALS- AB) ÷W
3 ) Bacteria or Zellen:
TG(U/104 cell) = 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: Sample protein concentration (mg/ml); W: Sample weight(G);
D: Density of bacteria or cell, 104 cell/mL.
Notiz:
- 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
- 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
- In order to ensure the repeatability of the test, the cooling time after each water bath should be
- 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]. 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.
Verwandte Produkte
BC1890/BC1895 Free Cholestenone (FC) Content Assay Kit
BC0750/BC0755 Acetaldehyde Dehydrogenase(ALDH) Aktivitätstest-Kit
BC0410/BC0415 Acetyl CoA carboxylase(ACC) Aktivitätstest-Kit
BC1980/BC1985 Total Cholesterol(TC) Content Assay Kit
Technische Spezifikationen:
The detection limit: 0.0372 mg/ml
Linear range: 0.0625-3 mg/ml
Bewertungen
Es gibt noch keine Bewertungen.