Solarbio Tissue Total Phosphorus Content Assay Kit

Tissue Total Phosphorus Content Assay Kit

Catatan: Ambil dua atau tiga sampel berbeda untuk prediksi sebelum pengujian.

Detection instrument: Spectrophotometer /Microplate reader

Kucing No: BC2855

Ukuran: 100T/96S

Komponen:

Reagen I: 20 mL×1. Penyimpanan pada suhu 4℃ (strong corrosive, strong oxidizing). Reagen Ⅱ: 7 mL×1. Penyimpanan pada suhu 4℃.

Reagen III: Bedak×2. Storage at 4℃ and protected from light. Solusi kerja: Fully dissolved with 5 mL air suling, lalu tambahkan 2.5 mL of Regent II, campur dengan baik.

Standar: 1 mL×1, 1 mmol/L inorganic phosphorus standard. Penyimpanan pada suhu 4℃.

Deskripsi Produk:

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, dan sebagainya. 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.

After digestion, 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

Mesin sentrifugal, Spectrophotometer/Microplate Reader, mandi air, kuvet kaca mikro/pelat dasar datar sumur 96, pipet transfer, air suling, concentrated sulfuric acid.

Prosedur:

SAYA. Ekstraksi Sampel:

0.1g of sample with 1mL of concentrated sulfuric acid (cover tightly to prevent moisture loss) put in boiling water bath for 10 menit. When the solution is black or brown, take it out. Add 200μL of Reagent I after cooling, campur dengan baik. 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 menit, Supernatan digunakan untuk tes.

II. Prosedur Penentuan:

1. Preheat the spectrophotometer/ microplate reader for 30 menit, sesuaikan panjang gelombang ke 660 nm, atur nol dengan air suling.
2. Set the temperature of water bath to 40℃.
3. Tambahkan reagen dengan daftar berikut:

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

Perhitungan:

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

=0.005×(PADA – AB)−(SEBAGAI – AB)−W

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

Catatan:

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.

Contoh eksperimental:

1. Take 0.1g kidney according to the extraction procedure, centrifugally take it up and clean it, and then follow the measurement procedure. Menggunakan 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. Menggunakan 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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Spesifikasi teknis:

Batas deteksi: 0.0338 mmol/L Linear range: 0.625-8 mmol/L