Anti HA Antibody Beads

Anti HA Antibody Beads is the slurry of anti HA monoclonal antibody immobilization beads. This product is used for the purification of HA tagged recombinant proteins by the competitive elution using HA peptide (YPYDVPDYA).

[Formulation]
1 x PBS (pH 7.4), 50% glycerol, 0.02 w/v% sodium azide.

[Beads matrix]
4% Agarose

[Antibody quantity]
8.5 mg/mL beads

[Antibody clone No.]
4B2

[Antibody subclass]
IgG₁

[Binding capacity]
Min. 1.5 mg HA tagged recombinant protein/mL beads

[Setting Volume]
1.8 - 2.1 mL slurry/mL beads

[Additional materials required]
centrifuge, centrifugation tube, 1.5 mL micro centrifugation tube, washing buffer (PBS(-), TBS etc.), cell scraper, protease inhibitor, phosphatase inhibitor, cell lysis buffer (following).

Example of cell lysis buffer
- 50 mmol/L Tris-HCl pH 7.4, 15 mmol/L Sodium Chloride, 1 mmol/L EDTA, 1% Triton X-100.
- RIPA Buffer (Code No. 182-02451)
50 mmol/L Tris-HCl, pH 8.0, 150 mmol/L Sodium Chloride, 0.5 w/v% Sodium Deoxycholate, 0.1 w/v% Sodium Dodecyl Sulfate, 1.0 w/v% NP-40 substitute.
- Cell Lysis Buffer M (Code No. 038-21141)
20 mmol/L Tris-HCl, pH 7.4, 200 mmol/L Sodium Chloride, 2.5 mmol/L Magnesium Chloride, 0.05 w/v% NP-40 substitute.

[Procedure]
The immunoprecipitation of HA tagged recombinant proteins (Mammalian cells).

1. The preparation of cell lysates
A. Adherent cells
B. Suspension cells

2. Antigen-antibody reaction

3. The elution of HA tagged recombinant proteins
A. The competitive peptide elution at neutral pH condition by using HA peptide
B. The elution at acidic condition by using 0.1 mol/L Glycine-HCl (pH 3.5) (room temperature)
C. The elution by using SDS sample buffer (room temperature)

- Each procedure is shown on the package insert.