Usage (IgG as an example, for reference only)

IgG reduction

For better labeling, the antibody solution is recommended at 1 mg/mL, and if it is lower than 1 mg/mL, the antibody should be concentrated. Antibody reduction can be performed in phosphate buffers. The type, concentration and incubation time of reducing agent can refer to the corresponding literature. After purification, the concentration of the reducing antibody should be determined.

Note:

Reducing antibodies are unstable and it is best to conduct the coupling reaction immediately after purification.

Marking steps (for reference only)

1. The reduced antibody is added to the pre-activated dye tube at the ratio of 1.5:1 (mass ratio).

Note:

The pre-activated PE-Cy dye 1 mg specification is recommended for one-time use, that is, if the antibody solution concentration is 1 mg/mL, according to the mass ratio of 1.5:1, 667 μL of antibody solution is required to be added to the pre-activated dye.

2. Shake and incubate at room temperature (20-25℃) for 4 h away from light.

3. Block the free sulfhydryl group on the antibody, need to refer to the literature.

4. Use volume exclusion chromatography to purify antibody /PE-Cy conjugate solution.

Note:

If long-term storage, need to add antibody preservation solution.

Matters needing attention

1. To label different antibodies, the required dye and antibody labeling ratio may be different. When the antibody quality, labeling steps and other factors that may affect the labeling result are excluded, the antibody quality can be reduced to achieve a better labeling effect.

2. Fluorescent dyes are all quenched, please pay attention to avoid light to slow down fluorescence quenching.

3. For your safety and health, please wear a lab coat and disposable gloves.