Alkali-tolerant Protein A Sepharose FF (rat-PA Sepharose FF)
Product Name: Alkali-tolerant Protein A Sepharose FF (rat-PA Sepharose FF)
Catalog Number: NRPB06L, NRPB06S (prepacked column)
Packing Details: 10 ml, 100 ml, 1 L, 1 ml (prepacked column), 2 ml (prepacked column), 5ml (prepacked column), 10ml (prepacked column)
Introduction:
Protein A is a cell wall protein of Staphylococcus aureus and contains five functional domains that can bind to the Fc fragment of IgG. Structural modification of nature protein A improved its alkaline resistance. Therefore, NaOH could be used to regenerate resin. A design that spacer arms on sepharose backbone are conjugated with the oriented binding sites of recombinant alkali-tolerant protein A ensures a formation of strong and efficient conjugate between sepharose and recombinant alkali-tolerant protein A as well as a higher capacity of recombinant alkali-tolerant protein A than native protein A.
Column Characteristics:
Support 6% highly cross-linked spherical agarose
Ligand rat-PA
Ligand Density (mg/ml) ＞6
Particle Size (μm) 45~165
Flow Rate-Recommended/Maximum (cm/h) 100/750
pH Limits-Working/Cleaning 3~11/2~13
Maximum Operating Pressure (MPa) 0.3
Capacity ≥35 mg/ml IgG
Storage 4 oC to 8 oC in 20% ethanol
Performing A Separation:
Binding buffer: 20 mM sodium phosphate, 150 mM NaCl, pH 7.4
Elution buffer: 100 mM sodium citrate, pH 4.0
Neutralization buffer: 1M Tris-HCl, pH 9.0
1) Wash the prepacked 1 ml column with 5~10 column volumes of distilled water to remove 20% ethanol.
2) Equilibrate the column with 5~10 column volumes of binding buffer.
3) Dilute 5 ml rabbit serum with binding buffer to 50 ml. Filtrate the diluted serum through a 0.45 μm filter and load the sample.
4) Wash with 10 column volumes of binding buffer.
5) Elute with 5 column volumes of elution buffer and neutralize collect fractions with neutralization buffer.
6) After each separation cycle, regenerate the resin by washing with approximately 3~5 column volumes of 0.1 M NaOH (pH 3.0).
7）Confirm the purity of the collected antibody by SDS-PAGE analysis (>95% purity).
