Aug 11, 2020
Purification of anti-anti-SpA antibody (Ab-2) using SpA-bearing Staphylococcus aureus cells.
- 1University of the West Indies St. Augustine
- University of the West Indies
- angel.vaillant@sta.uwi.edu
Protocol Citation: Angel A Justiz-Vaillant 2020. Purification of anti-anti-SpA antibody (Ab-2) using SpA-bearing Staphylococcus aureus cells.. protocols.io https://dx.doi.org/10.17504/protocols.io.bjj9kkr6
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Created: August 11, 2020
Last Modified: August 11, 2020
Protocol Integer ID: 40289
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Abstract
A novel method was designed for the purification of Ab-2 from undiluted WSF of hyperimmune eggs. A total of six preparations were made.
Materials
MATERIALS
Centrifuge 5424 R refrigerated with Rotor FA-45-24-11 rotary knobs 120 V/50 – 60 Hz (US)Eppendorf CentrifugeCatalog #5404000537
Eppendorf Safe-Lock Tubes 1.5 mL PCR clean colorless 500 tubesEppendorfCatalog #022363212
SpA-bearing Staphylococcus aureus cells.
Mix in an Eppendorf microtube 0.9 ml of undiluted WSF from immunized birds with 25 μl of SpA-bearing Staphylococcus aureus cells (Sigma-Aldrich).
Incubate the microtube at 37oC for 30 min.
After the incubation period, centrifuge the microtube in an Eppendorf 5424 centrifuge for 5 min.
Observe the microtube that presents a pellet of Ab-1 bound-cells at the bottom ,and the supernatant, where Ab-2 is present.
Decant the Ab-2 containing supernatant to another tube.
. Store it at -20oC until further use.