Protein A (ProA, Staphylococcus A Protein) (AP)

Molecular Biology / MB-BioSep Protein A/G/L

4°C Do Not Freeze

Supplied as a liquid in 0.05M tris chloride, 0.15M sodium chloride, 0.001M magnesium chloride, 0.0001M zinc chloride, pH 8.0, 0.09% sodium azide, 50% glycerol, 10mg/ml BSA. Labeled with Alkaline Phophatase (AP).

Protein A is a surface protein (approximately 56kD in size) originally discovered within the cell wall of Staphylococcus aureus. While important for bacterial survival, Protein A has beneficial uses in immunology for its high affinity binding to immunoglobulins (especially the IgG isotype). This high affinity property makes Protein A essential in the large scale purification. Alkaline Phosphatase is an enzyme which removes phosphate groups from a variety of substate molecules. As the name implies, this enzyme functions best under basic pH. Alkaline Phosphatase can be utilized in molecular biology in DNA ligation experiments (keeping the DNA linear), radiolabeling preparations, and a detection mediator in ELISA experiments.

Source:
Protein A Alkaline Phosphatase Conjugated was prepared from chromatographically pure Protein A. Protein A Alkaline Phosphatase Conjugated assayed by immunoelectrophoresis resulted in a single precipitin arc against anti-Alkaline Phosphatase (calf intestine) and anti-Protein A. No reaction was observed against anti-Protein G.

Applications:
Suitable for use in ELISA, Western Blot and Immunohistochemistry. Other applications not tested.

Recommended Dilution:
ELISA: 1:8000-1:32, 000
Western Blot: 1:500-1:2500
Immunohistochemistry: 1:200-1:1000
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4°C before opening. DO NOT FREEZE! Stable at 4°C as an undiluted liquid. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. Freezing alkaline phosphatase conjugates will result in a substantial loss of enzymatic activity.