Catatan: Ambil dua atau tiga sampel berbeda untuk prediksi sebelum pengujian.
Peralatan Operasi: Spectrophotometer/Microplate Reader
Nomor Katalog: BC5175
Ukuran: 50T | 48S
Komponen:
Reagen I: 25 mL×1. Store at 2-8℃.
Reagen II: 6 mL×1. Store at 2-8℃.
Deskripsi Produk：
Bilirubin langsung (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.
Reagen dan Peralatan Diperlukan tetapi Tidak Disediakan：
Spectrophotometer/Microplate Reader, mesin sentrifugal meja, constant temperature foster box/water bath, pipette, kuvet kaca mikro/pelat dasar datar sumur 96, Es, dan air suling.
Prosedur
A. Persiapan sampel:
Serum, plasma, or other liquid samples: Detect the sample directly. If the solution is turbid, measure centrifuging.
B.Tekad prosedur：
- Preheat the spectrophotometer/microplate reader for 30min, sesuaikan panjang gelombangnya 450 nm, and set the spectrophotometer counter to zero with distilled
- Tekad:
| Reagen (μL) | Tabung reaksi | Tabung kosong |
| Sampel | 8 | – |
| Air sulingan | – | 8 |
| Reagen I | 192 | 192 |
| Aduk rata. React at 37℃ for 5 menit. Ukur serapannya pada 450 nm, and record it as A1T, A1B; | ||
| Reagen II | 48 | 48 |
| Aduk rata. React at 37℃ for 5 menit. Ukur serapannya pada 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.Perhitungan：
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
Catatan:
- 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.
Contoh:
- 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.
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Ulasan
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