Anti DYKDDDDK tag Antibody Beads

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

[Beads matrix]
4% Agarose

[Antibody quantity]
Approx. 7.5 mg/mL beads

[Antibody clone No.]
1E6

[Antibody subclass]
IgG_2b

[Binding capacity]
Min. 1.0 mg 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, 150 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 DYKDDDDK tagged rProteins (Mammalian cells).

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

2. Antigen-antibody reaction

3. The elution of DYKDDDDK tagged rProtein
A. The competitive peptide elution at neutral pH condition by using DYKDDDDK 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.