1 Sample Pretreatment
Take a sample with an antibody and dilute it to 50 uL with the binding solution. If the sample volume is greater than 50 uL, no dilution is required.
2 Magnetic Beads Pretreatment
2.1. Dispersion: Vortex the magnetic beads, or shake it, or mix upside down to fully resuspend the magnetic beads;
2.2. Pipette magnetic beads and transfer them to a centrifuge tube;
2.3. Pre-Washing: add binding solution to the centrifuge tube, mix well, place the centrifuge tube on a magnetic stand, and let it stand to make all the magnetic beads adsorb on the tube wall. The supernatant is clear and transparent. Pipette Aspirate and discard the supernatant.
2.4. Repeat the pre-wash step twice. After washing, the supernatant magnetic beads should be used as soon as possible to prevent drying and aggregation.
3 Antibody Binding
3.1. Add the pre-processed sample to the pre-processed magnetic beads, pipette gently to mix and avoid foam;
3.2. Incubate by turning the mixer or manually gently turning At room temperature;
3.3. Magnetic separation: Place the centrifuge tube on a magnetic stand and let it stand to make all the magnetic beads adsorb on the tube wall. Pipette and discard the supernatant.
3.4. Add binding solution to the centrifuge tube, mix well, place the centrifuge tube on a magnetic stand, magnetically separate, and discard the supernatant;
3.5. Add binding solution to the centrifuge tube, mix thoroughly, transfer to a clean centrifuge tube, magnetically separate, and discard the supernatant.
4 Antibody Elution
4.1. Add eluent to the centrifuge tube that has completed the antibody binding step, pipette gently to mix, and try to avoid foaming.
4.2. Incubate by turning over the mixer or manually gently turning over At room temperature
4.3. Magnetic separation, carefully aspirate the supernatant with a pipette, and transfer to a clean centrifuge tube.
4.4. Immediately add neutralization solution to the eluted supernatant, and mix gently.
Note: This section provides a general antibody purification procedure. Different types of antibodies of different quality can be optimized on this basis to obtain the best purification results.