Protein A Beads

Details for Product No. ABIN921275, Supplier: Log in to see
  • alpha protein
  • repB
Staphylococcus aureus (S. aureus)
Separation (Sep), Purification (Purif)
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Purpose Protein A Beads for affinity purification of antibodies.
Specificity Protein A binds to most human and mouse IgG subclasses (e.g. human IgG1, IgG2, IgG3, IgG4, IgA, mouse IgG1, IgG2a, IgG2b, IgG3). It also binds to rat IgG1, IgG2c, goat IgG1, IgG2, sheep IgG1, IgG2, cow IgG1, IgG2, horse IgG (ab), IgG(c). Protein A binds strongly to total IgG from rabbit, dog, cat, pig, guinea pig.
Characteristics Protein A Beads Specifications
Matrix: CNBr-activated SepharoseTM 4FF.
Beads concentration: 1-2 mg/ml.
Coupling conditions of matrix: pH 7-9, 4°C to 25°C, 2-16 h.
Binding capacity: 4-7 mg IgG per ml.
Bead size range: 45-165 m.
Mean bead size: 90 m.
Bead structure: Highly cross-linked agarose, 4%.
Max. flow rate: 4 ml/min/cm2.
Recommended flow rate: 1-3 ml/min/cm2.
Stability of the matrix: pH 3-11 (ligand dependent).
Bead Ligand Protein A
Bead Matrix Sepharose beads
Bead Size 90 µm
Application Notes This product can be used for 100-200 times. For frequent use, an aliquot can be stored at 4ºC for 1 month with addition of 0.02% sodium azide (NaN3) to the storage buffer. Because this product can purify IgG subclasses from several species of mammals.
Reagent Preparation

Buffers preparation: Equilibration buffer A: 1% Nacl+0.1% Na 2HPO4, pH7.5. Equilibration buffer B: 1% CH3COONa adjusted pH to 5 by CH3COOH. Elution buffer: 2% table sugar adjusted pH to 2-3 by CH3COOH. Wash buffer: purified water. Storage buffer: 30% glycerol.

Sample Preparation

Sample preparation: 1. Dilute the serum with equilibration buffer A to ensure its content and pH closed to equilibration buffer A. 2. Centrifuge diluted serum supernatants to sediment debris. 3. Filter supernatants through 0.45m filter.

Assay Procedure

Affinity-purification: 1. Load the Protein A beads into the empty column. 2. Wash column with Wash buffer in 3-5 column volumes to remove the glycerol, and then, equilibrate column by washing with Equilibration buffer A in 5-10 column volumes. 3. Bring the sample to room temperature, and load it into the column by a syringe or a pump. The total volume of the sample applied is not critical in most cases. 4. Load the sample into the column and collect the flow liquid, repeat this action for 3-5 times. If necessary, repeat for more times, then deal with the collected liquid reasonably. 5. Wash the column with Equilibration buffer B to remove other proteins. 6. Elute with Elution buffer, collect the flow liquid (antibody), adjust its pH by saturated Na2CO3 during collection. Then, customers can test the related data of the antibody as their own requirements. Re-equilibration and Storage: 1. Add 5-10ml Elution buffer to column to elute thoroughly, then neutralizate the column with Equilibration buffer A. 2. Wash the column bed with Storage buffer in 3-5 column volumes, seal the bottom of the column and store at -20°C for at least one year. For frequent use, an aliquot can be stored at 4°C for 1 month with addition of 0.02% sodium azide (NaN 3) to the storage buffer.

Restrictions For Research Use only
Format Liquid
Concentration 1-2 mg/mL
Buffer Equilibration buffer A: 1% NaCl+0.1% Na2HPO4 , pH 7.5
Equilibration buffer B: 1% CH3COONa adjusted pH to 5 by CH3COOH
Elution buffer: CH3COOH (pH 2-3) or 0.1mol Glycine Hydrochlorde.
Wash buffer: 1% NaCl+0.1% Na2HPO4, pH 7.5
Storage buffer: 30% glycerol
Handling Advice Note: 20% ethanol was contained as protection solution in this product, please wipe off the ethanol before use.
Storage 4 °C/-20 °C
Storage Comment At -20°C for at least one year. Store at 4°C for frequent use.
Background publications Yu, Fang, Zhu, Wang, Tien, Zhang, Chen: "One time intranasal vaccination with a modified vaccinia Tiantan strain MVTT(ZCI) protects animals against pathogenic viral challenge." in: Vaccine, Vol. 28, Issue 9, pp. 2088-96, 2010 (PubMed).