Solarbio Tissue Total Phosphorus Content Assay Kit

Tissue Total Phosphorus Content Assay Kit

Nota: Tome dos o tres muestras diferentes para predecir antes de la prueba..

Instrumento de detección: Spectrophotometer /Microplate reader

No gato: BC2855

Tamaño: 100T/96S

Componentes:

reactivo yo: 20 mL×1. Almacenamiento a 4 ℃ (strong corrosive, strong oxidizing). Reactivo ⅱ: 7 mL×1. Almacenamiento a 4 ℃.

Reactivo III: Polvo×2. Storage at 4℃ and protected from light. Solución de trabajo: Fully dissolved with 5 ml de agua destilada, Luego añade 2.5 mL of Regent II, mezclar bien.

Estándar: 1 mL×1, 1 mmol/L inorganic phosphorus standard. Almacenamiento a 4 ℃.

Descripción del Producto:

The form of phosphorus includes inorganic phosphorus and organic phosphorus. Inorganic phosphorus mainly refers to phosphate radical, which is involved in many kinds of metabolism, including energy metabolism, nucleic acid metabolism, protein phosphorylation, dephosphorylation, etcétera. By measuring the content of total phosphorus and inorganic phosphorus, the utilization rate of phosphorus in crops can be understood, and the basis for rational fertilization can be provided.

Después de la digestión, total phosphorus was converted into inorganic phosphorus. The molybdenum blue method is a classical method for determining the content of inorganic phosphorus. Under certain conditions, molybdenum blue and phosphate from a substance with a characteristic absorption peak at 660 Nuevo Méjico. By measuring the light absorption of 660 Nuevo Méjico, the inorganic phosphorus content can be calculated, and then the total phosphorus content in the tissue can be calculated.

Material requerido

Centrífugo, lector de espectrofotómetro/microplato, baño de agua, cubeta de microvidrio/placa de fondo plano de 96 pocillos, pipeta de transferencia, agua destilada, ácido sulfúrico concentrado.

Procedimiento:

I. Extracción de muestras:

0.1g of sample with 1mL of concentrated sulfuric acid (cover tightly to prevent moisture loss) put in boiling water bath for 10 minutos. When the solution is black or brown, take it out. Add 200μL of Reagent I after cooling, mezclar bien. Continue boiling (cover tightly) until the solution is transparent, then cool to room temperature, add 3.8mL of distilled water and mix well. Centrifugado en, 10000rpm and room temperature for 10 minutos, supernatant is used for test.

II. Procedimiento de determinación:

1. Preheat the spectrophotometer/ microplate reader for 30 mín., ajustar la longitud de onda a 660 Nuevo Méjico, poner a cero con agua destilada.
2. Set the temperature of water bath to 40℃.
3. Agregue reactivos con la siguiente lista:

Mezclar bien, 40℃ water bath for 10 minutos, detect the absorbance at 660 nm after cooling at room temperature for 10 minutos. Record as AB, AS and AT respectively.

Cálculo:

Total phosphate (mmol/g fresh weight)=[C×(EN- AB)÷(COMO – AB)]×V÷W

=0.005×(EN – AB)÷(COMO – AB)÷W

C: standard concentration, 1mmol/L; V: supernatant volume, 5 mL=0.005 L; W.: Peso de la muestra, gramo.

Nota:

1．Reagent III should be prepared before use and only be used the same day.

2．Before the determination, 1–2 samples shall be used for pre-test. If the absorption value is greater than 1.5, distilled water shall be used for corresponding dilution.

Ejemplo experimental:

1. Take 0.1g kidney according to the extraction procedure, centrifugally take it up and clean it, and then follow the measurement procedure. Usar 96 well plate to calculate: AT= 0.191, AB= 0.051, AS= 0.282. Calculate the total phosphorus content according to the sample mass

Total phosphorus content (mmol/g mass) = 0.005 × (A-AB) ÷ (AS-AB) ÷ W= 0.005×(0.191-0.051) ÷ (0.282-0.051) ÷ 0.1= 0.030 mmol/g mass.

2. Take 0.1g spleen according to the extraction procedure, centrifugally take the cleaning, and then follow the measurement procedure. Usar 96 well plate to calculate: AT= 0.197, AB= 0.051, AS= 0.282, calculate the total phosphorus content according to the sample mass

Total phosphorus content (mmol/g mass) =0.005×(A-AB) ÷(AS-AB)÷W= 0.005 × (0.197-0.051) ÷ (0.282- 0.051) ÷ 0.1= 0.032 mmol/g mass.

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Especificaciones técnicas:

El límite de detección: 0.0338 mmol/L Linear range: 0.625-8 mmol/L