Tissue Total Phosphorus Content Assay Kit
Nota: Prendi due o tre campioni diversi per la previsione prima del test.
Strumento di rilevamento: Spectrophotometer /Microplate reader
Gatto n: BC2855
Misurare: 100T/96S
Componenti:
Reagente I: 20 ml×1. Conservazione a 4 ℃ (strong corrosive, strong oxidizing). Reagente Ⅱ: 7 ml×1. Conservazione a 4 ℃.
Reagente III: Polvere×2. Storage at 4℃ and protected from light. Soluzione funzionante: Fully dissolved with 5 mL di acqua distillata, quindi aggiungi 2.5 mL of Regent II, mescolare bene.
Standard: 1 ml×1, 1 mmol/L inorganic phosphorus standard. Conservazione a 4 ℃.
Descrizione del prodotto:
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, e così via. 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.
Dopo la digestione, 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.
Materiale richiesto
Centrifuga, Spettrofotometro/lettore di micropiastre, bagnomaria, microcuvetta in vetro/piastra a fondo piatto da 96 pozzetti, pipetta di trasferimento, acqua distillata, concentrated sulfuric acid.
Procedura:
IO. Estrazione del campione:
0.1g of sample with 1mL of concentrated sulfuric acid (cover tightly to prevent moisture loss) put in boiling water bath for 10 minuti. When the solution is black or brown, take it out. Add 200μL of Reagent I after cooling, mescolare bene. 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 minuti, Il surnatante viene utilizzato per il test.
II. Procedura di determinazione:
- Preheat the spectrophotometer/ microplate reader for 30 min, regolare la lunghezza d'onda su 660 nm, azzerare con acqua distillata.
- Set the temperature of water bath to 40℃.
- Aggiungere i reagenti con il seguente elenco:
| Nome del reagente (μL) | Tubo vuoto (AB) | Tubo standard (A) | Provetta (COME) |
| Standard | 10 | ||
| Supernatante | 10 | ||
| Acqua distillata | 100 | 90 | 90 |
| Reagente III | 100 | 100 | 100 |
Mescolare bene, 40℃ water bath for 10 minuti, detect the absorbance at 660 nm after cooling at room temperature for 10 minuti. Record as AB, AS and AT respectively.
Calcolo:
Total phosphate (mmol/g fresh weight)=[C×(A- AB)÷(COME – AB)]×V÷W
=0.005×(A – AB)÷(COME – AB)÷W
C: standard concentration, 1mmol/l; V: supernatant volume, 5 mL=0.005 L; W: Peso del campione, G.
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.
Esempio sperimentale:
- Take 0.1g kidney according to the extraction procedure, centrifugally take it up and clean it, and then follow the measurement procedure. Utilizzo 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.
- Take 0.1g spleen according to the extraction procedure, centrifugally take the cleaning, and then follow the measurement procedure. Utilizzo 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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Specifiche tecniche:
Il limite di rilevamento: 0.0338 mmol/L Linear range: 0.625-8 mmol/l
Recensioni
Non ci sono ancora recensioni.