HA Tag Immunomagnetic Beads

• Low non-specific binding
• Time saving and efficient usage
• Convenient and simple operation
• Assay consistency

1. Preparation of cell lysates

Select an appropriate lysis buffer to lyse cell samples and obtain cell lysates. Place on ice or store at -20℃ for long-term use.

2. Pretreatment of Magnetic Beads

(1) Vortex for 1 min to resuspend the immunomagnetic beads. Take 25-50 µL of suspension and place it in a 1.5 mL EP tube.

(2) Add 500 μL of Washing Buffer to the EP tube and gently invert several times to resuspend the beads. Keep the EP tube in a magnetic separator and stand for 1 min for magnetic separation. Finally, remove the supernatant and then take off the EP tube. Repeat the washing steps twice.

3. Immunoprecipitation

(1) Add 500 μL of prepared cell lysates to the EP tube. Place it on a rotating mixer and rotate at 37℃ for 30 min. For weak binding, incubate at room temperature for 1 hour or overnight at 4℃.

(2) After incubation, perform magnetic separation, then remove or save the supernatant for further analysis.

(3) Add 500 μL of Washing Buffer to the EP tube. Perform magnetic separation. Finally, remove the supernatant and then take off the EP tube. Repeat the washing steps 3 times.

4. Elution of Target Proteins

(1) Denaturing Elution: Suitable for SDS-PAGE detection. Add 100 μL of SDS-PAGE Loading Buffer to the EP tube. Mix well and heat at 95℃ for 5 min. Perform magnetic separation or centrifugation (room temperature, 13000 g, 10 min) to collect the supernatant.

(2) Neutral Elution: Add 50 μL of HA Peptide Elution Buffer to the EP tube. Incubate on a rotating mixer at 37℃ for 5-10 min (longer incubation time when below 37℃). Then perform magnetic separation or centrifugation to collect the supernatant.

(3) Acidity Elution: Add 100 μL of Acidity Elution Buffer to the EP tube. Incubate on a rotating mixer at 37℃ for 5-10 min. Perform magnetic separation or centrifugation to collect the supernatant. To adjust the pH of acidic elution buffer to neutral, add 50 μL of Neutralization Buffer to 100 μL elution.

Store at 4℃ for 2 years.

1. Avoid freezing the beads. Store in solution to prevent drying.
2. The average magnetic separation time should be longer than 1 min.
3. Ensure uniform suspension by fully shaking the storage tube before use. Avoid bubbles during operation.
4. Use high-quality tips and test tubes to avoid sample loss due to adhesion.
5. Test the binding of proteins to beads by using the collected supernatant.
6. In IP experiments, the binding affinity of different proteins may vary. Users can select and prepare buffers according to experimental needs.
7. The product is for R&D use only, not for diagnostic procedures, food, drug, household or other uses.
8. Please wear a lab coat and disposable gloves.