Biotin, a small molecule with a molecular weight of only 244.31 Da, can be combined with biological macromolecules such as proteins and antibodies after activation. It not only maintains the biological activity of macromolecules well, but also has multiple levels when used in combination with avidin. Amplification, it is widely used in micro-antigen, antibody qualitative, quantitative detection and localization observation research. So how do you use biotin to label antibodies or proteins? Let me share it for you. Tools / Materials • 10ul, 50ul, 200ul, 1000ul adjustable high precision pipette • Thermostat (37°C) Preparation before experiment Operation steps: (This procedure is marked according to the amount of 1mg) 2. Add 13.3 ul of NH2-Reactive Biotin and the appropriate amount of Labeling Buffer to the above Filtration tube to a final volume of 0.5 ml and mix gently by pipetting. Incubate in a 37 ° C incubator for 30 min in the dark. Centrifuge at 12,000 xg for 10 min. 5 Inch Face Access Control,5-Inch Face Recognition Attendance Access Control,Face Recognition Attendance Access Control,Office Face Recognition Access Control System Shenzhen BIO Technology Co.,Ltd. , https://www.hfsecuritytech.com
• Centrifuge (centrifugal force up to 12,000 × g)
• Biotin Labeling Kit (Cata.No: EBLK0002, Elabscience)
1. Read the instruction manual carefully.
2. Calculate the amount of NH2-Reactive Biotin to be used.
3. Remove the kit from the refrigerator 20 minutes in advance and equilibrate to room temperature (Note: NH2-Reactive Biotin does not need to be used in the refrigerator).
4. Dissolve NH2-Reactive Biotin: Add 30 ul of DMF to NH2-Reactive Biotin bottle and let stand for 10 min until it is fully dissolved. At this time, the concentration of biotin is 10 mM.
1. Take 1 mg of the antibody to be labeled in a Filtration tube, and add a corresponding volume of Labeling Buffer to make the final concentration of the antibody 2 mg/ml and centrifuge at 12,000 xg for 10 min. (Note: 1Filtration tube has a maximum volume of 0.5ml2. When the concentration of the antibody to be labeled is low, it can be centrifuged once by ultrafiltration.)
3. Add the appropriate amount of Labeling Buffer to the above Filtration tube to a final volume of 0.5 ml, mix gently by pipetting, and centrifuge at 12,000 xg for 10 min.
4. Repeat 3 for operation 3.
5. Add 0.2ml Labeling Buffer to the Filtration tube and gently blow. The filter element was placed upside down in another centrifuge tube and centrifuged at 6,000 xg for 10 min.
6. Collect the solution in the centrifuge tube, which is a biotinylated antibody.