Notiz: Take two or three different samples for prediction before the test.
Betriebsausrüstung: Spectrophotometer/Microplate reader
Catalog Number: BC5175
Größe: 50T | 48S
Komponenten:
Reagenz I: 25 ml×1. Store at 2-8℃.
Reagenz II: 6 ml×1. Store at 2-8℃.
Produktbeschreibung：
Direct bilirubin (DBIL) is also called conjugated bilirubin. After indirect bilirubin enters the liver, it is combined with glucuronic acid by the action of glucuronosyltransferase in the liver. The increase of direct bilirubin is of great significance for clinical diagnosis of obstructive jaundice, hepatocellular jaundice, liver cancer, pancreatic head cancer, cholelithiasis, and cholangiocarcinoma. Direct bilirubin could be oxidized by sodium nitrite to form biliverdin, which has an absorbance of 450 nm. The content of direct bilirubin can be calculated by detecting the wavelength change at 450 nm.
Erforderliche, aber nicht bereitgestellte Reagenzien und Ausrüstung：
Spectrophotometer/Microplate reader, desk centrifuge, constant temperature foster box/water bath, pipette, Mikroglasküvette/96-Well-Platte mit flachem Boden, Eis, and distilled water.
Verfahren
A. Probenvorbereitung:
Serum, Plasma, or other liquid samples: Detect the sample directly. If the solution is turbid, measure centrifuging.
B.Bestimmung Verfahren：
- Preheat the spectrophotometer/microplate reader for 30min, adjust the wavelength to 450 nm, and set the spectrophotometer counter to zero with distilled
- Bestimmung:
| Reagens (μL) | Test tube | Blank tube |
| Probe | 8 | – |
| Distilled water | – | 8 |
| Reagenz I | 192 | 192 |
| Mix well. React at 37℃ for 5 Protokoll. Measure the absorbance at 450 nm, and record it as A1T, A1B; | ||
| Reagenz II | 48 | 48 |
| Mix well. React at 37℃ for 5 Protokoll. Measure the absorbance at 450 nm, and record it as A2T, A2B. Calculate the ΔAT =A1T-A2T and the ΔAB =A1B-A2B. Blank tubes only need to be measured once or twice. | ||
C.Berechnungen：
96 well flat-bottom plate
DBIL content (μmol/L) =1495.7×(ΔAT-ΔAB)-15.343
Micro glass cuvette
DBIL content (μmol/L) =1099.7×(ΔAT-ΔAB)-10.73
Notiz:
- Bilirubin decomposes easily in light. Avoid light during
- If the ΔA is lower, it is recommended to increase the sample size before determination; If ΔA＞5, it is recommended to dilute the sample before determination.
Examples:
- Take mouse serum to follow the determination procedure to operate. Determination with 96 well flat-bottom plate, and calculate ΔAT=A1T-A2T=0.217-0.170=0.047, ΔAB=A1B-A2B=0.044-0.044=0. The calculated content is as follows:
DBIL content (μmol/L) =1495.7×(ΔAT-ΔAB)-15.343=54.95 μmol/L.
Related products
BC5180/BC5185 Total bilirubin (TBIL) Content Assay Kit
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