Determine the necessary mass, volume, or concentration for preparing a solution.
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Basic Description
| Synonyms | Hematoidin Assay Kit | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Specifications & Purity | Colorimetry | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Storage Temp | Store at -20°C | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Shipped In |
Ice chest + Ice pads This product requires cold chain shipping. Ground and other economy services are not available. |
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| Grade | Colorimetry | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Product Description |
Bilirubin, also known as bilirubin like, is a degradation product formed by the breakdown and metabolism of hemoglobin in the liver. Bilirubin circulates in the bloodstream in an unbound insoluble form (indirect bilirubin) or a soluble glucoside bound form (direct bilirubin). Combined with bilirubin, it is transferred from the bile ducts of the liver to the gallbladder and secreted into the small intestine during digestion. Excessive levels of bilirubin can lead to jaundice and indicate possible liver disease, blood disorders, or bile duct obstruction.
The bilirubin detection kit is used to detect the concentration of total bilirubin in samples. Applicability It can be used for the determination of total bilirubin and conjugated bilirubin in serum and plasma samples. Principle This detection method is based on the Jendrassik Grof method, which uses bilirubin to react with diazotized p-aminobenzenesulfonic acid to generate a colorimetric product that can be detected at 530 nm, which is proportional to the bilirubin present in the sample. This detection kit can detect total bilirubin and conjugated bilirubin. Sodium nitrite oxidation method Measurement principle At a pH value of approximately 2.9, sodium nitrite can oxidize bilirubin to biliverdin. In the presence of surfactant Triton X100, both free bilirubin and conjugated bilirubin (direct bilirubin) will undergo oxidation, and the decrease in absorbance at 450nm is proportional to the total bilirubin concentration. Determination of significance Total Bilirubin (TBIL) is the sum of direct bilirubin and indirect bilirubin. The liver plays an important role in the metabolism of bilirubin. Matters needing attention Before formal testing, it is necessary to take 3-5 samples with significant expected differences for prediction; For your safety and health, please wear protective gear; Self provided instruments and supplies Enzyme reader, 96 well plate, adjustable pipette, centrifuge, distilled water. Sample requirements 1、 Liquid samples The clarified liquid can be directly detected. If it is turbid, centrifuge and take the supernatant for testing be careful: 1. Due to the photooxidation of bilirubin after exposure to sunlight or ultraviolet radiation, sampling should be done quickly and attention should be paid to avoiding light; 2. Store the sample in a dark place at -70 ℃ for a maximum of 3 months; Experimental preparation 1. Preheat the enzyme-linked immunosorbent assay (ELISA) reader for more than 30 minutes, set the temperature to 37 ℃, and adjust the wavelength to 450nm; 2. Thaw reagent one and reagent two to room temperature; 3. Standard products are thawed at low temperature and placed on ice; Standard products can be stored in a dark place at 4 ℃ for a maximum of 1 week; It is recommended to pack according to the amount used each time and store at -20 ℃ for freezing (repeated freezing and thawing are not allowed). After use as needed, place at 4 ℃ for thawing; Measurement operation 1. According to experimental needs, distilled water can be used to dilute the concentration of the standard substance, and the diluted concentration of the standard substance can be calculated by substituting it into the formula (it is recommended to use the diluted standard substance on site or use it on the same day) 2. In a low light environment, operate sequentially in a 96 well plate
1. Bilirubin is easily decomposed when exposed to light, so be careful to avoid light during operation; 2. When using an enzyme-linked immunosorbent assay (ELISA) reader, pay attention to observe that there are no bubbles inside the 96 well plate; 3. If the Δ A value is less than 0.005, the sample volume can be increased appropriately and the reagent volume can be reduced accordingly; The following Methods can be used to change the sampling system:
Result calculation (1) Calculated by volume Total bilirubin content (μ mol/L)=(C × V2) x (△ A measurement - △ A blank) ÷ (△ A standard - △ A blank) ÷ V1 × D =C × (△ A measurement - △ A blank) ÷ (△ A standard - △ A blank) × D C: Standard concentration, 171 μ mol/L; V1: Add sample volume, 0.01mL; V2: Add standard sample volume, 0.01mL; D: Additional dilution factor, undiluted equals 1; The significance of pre experiments The preliminary experiment of colorimetric detection reagent kit is very important 1. Determine whether the reagent kit is suitable for customer sample testing to avoid wasting the reagent kit and samples (such as low expression treated samples); 2. Familiar with the operation process of biochemical reagent kits, especially for first-time use of biochemical reagent kits for testing; 3. Determine whether the sample processing method and dilution ratio are appropriate; 4. Understand possible experimental phenomena or issues that may arise during the experimental process, in order to make timely adjustments; 5. Through 3-5 pre experiments, determine the optimal dilution concentration range of the reagent kit for the sample and guide the dilution ratio of the experimental sample. |
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