Solarbio Tissue Total Phosphorus Content Assay Kit

Tissue Total Phosphorus Content Assay Kit

Notiz: Nehmen Sie vor dem Test zwei oder drei verschiedene Proben zur Vorhersage.

Erkennungsinstrument: Spectrophotometer /Microplate reader

Katze nein: BC2855

Größe: 100T/96S

Komponenten:

Reagenz I: 20 ml×1. Lagerung bei 4℃ (strong corrosive, strong oxidizing). Reagenz ⅱ: 7 ml×1. Lagerung bei 4℃.

Reagenz III: Pulver×2. Storage at 4℃ and protected from light. Funktionierende Lösung: Fully dissolved with 5 ml destilliertes Wasser, dann hinzufügen 2.5 mL of Regent II, gut mischen.

Standard: 1 ml×1, 1 mmol/L inorganic phosphorus standard. Lagerung bei 4℃.

Produktbeschreibung:

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, und so weiter. 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.

Nach Verdauung, 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 nm. By measuring the light absorption of 660 nm, the inorganic phosphorus content can be calculated, and then the total phosphorus content in the tissue can be calculated.

Required material

Zentrifuge, Spektrophotometer/Mikroplattenleser, Wasserbad, Mikroglasküvette/96-Well-Platte mit flachem Boden, Pipette übertragen, destilliertes Wasser, concentrated sulfuric acid.

Verfahren:

ICH. Probenextraktion:

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

II. Bestimmungsverfahren:

1. Preheat the spectrophotometer/ microplate reader for 30 Mindest, Wellenlänge anpassen 660 nm, Mit destilliertem Wasser auf Null stellen.
2. Set the temperature of water bath to 40℃.
3. Fügen Sie Reagenzien mit der folgenden Liste hinzu:

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

Berechnung:

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

=0.005×(BEI – AB)÷(ALS – AB)÷ w

C: standard concentration, 1mmol/L; V: supernatant volume, 5 mL=0.005 L; W: Probengewicht, G.

Notiz:

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.

Experimentelles Beispiel:

1. Take 0.1g kidney according to the extraction procedure, centrifugally take it up and clean it, and then follow the measurement procedure. Verwenden 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. Verwenden 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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Technische Spezifikationen:

The detection limit: 0.0338 mmol/L Linear range: 0.625-8 mmol/L