Immunoaffinity Column for Biotin

Description

1. Overview:

Biotin, also called vitamin B7, is one of the B vitamins. It is involved in a wide range of metabolic processes, both in humans and in other organisms, primarily related to the utilization of fats, carbohydrates, and amino acids.

2. Principle:

The basis of the measurement is the antigen-antibody reaction. Antibodies are connected to the column and the biotin in the sample is extracted, filtered, and diluted, and then passed slowly through the immunoaffinity column. The biotin binds to the antibodies in the column and the immunoaffinity column is then washed to remove other unrelated substances that have not been bound. Biotin is then eluted with methanol and injected into an analytical instrument for detection.

3. Components of the kit:

Each kit contains biotin immunoaffinity columns and 1 instruction manual.

4. Necessary items not provided in the box:

4.1 Equipment

• HPLC /LC-MS（LC-MS/MS）
• Nitrogen gas evaporatorapparatus
• Nitrogen gas tank and pressureregulator
• Air-pressure controller bracket
• Air pump
• Balance with 0.01greadability
• Ultrasoniccleaner
• High-speed freezing centrifuge (can be set to 10℃, 6000RPM)
• Centrifugetube:50-mL
• Constant temperature incubator or water bath (can be set to 35–37℃)
• High-speed homogenizer (maximum speed> 10,000 RPM) or shaker
• Grinder
• pH meter (or pH testpaper)
• Volumetric flask: 100 mL/50mL
• Syringe: 10 mL/20mL
• Homogenization flask (or 250-mL conical flask withpestle)
• Vials andtubes
• Column holder and syringe connector plug (for use withimmunoaffinity columns)

4.2 Reagents

• Methanol (CH3OH): Chromatography Grade
• Sodium hydroxide (NaOH): Analytical Grade
• Sodium chloride (NaCl): Analytical Grade
• Hydrochloric acid (HCl): Chromatography Grade
• Potassium dihydrogen phosphate (KH2PO4): Analytical Grade
• Dipotassium phosphate (K2HPO4): Analytical Grade
• Distilled or deionized water

5. Note:

• Allow the immunoaffinity column to return to room temperature (22 to 25°C) before use.
• The affinity column should be stored at 2 to 8°C, do not freeze.
• Dot use any expired immunoaffinity column.
• Column capacity:1000 ng, when the content of the biotin in the sample divided by the dilution factor is higher than the column capacity, it is necessary to reduce the volume of the sample solution appropriately, and retest.
• The pH of the loading solution of the affinity column should be between 6 to 8.

Keeping the solvent and mobile phase consistent with the instrument detection can eliminate the effect of solvent effects.