Description:
Micro Spin Filter provides an easy and efficient column method for Immunoprecipitation (IP), Co-IP, and Immunodepletion.
All the procedures involve 1) incubation of an antibody with a sample that contains the protein antigen of interest, 2)
capturing the antibody-antigen complex to immobilized Protein A or Protein G agarose beads, 3) washing to remove unbound
components of the sample, and 4) separation of antigen and antibody from beads.
Advantage of using spin filter column
Traditional method: The entire procedure is performed in a microcentrifuge tube, which requires a) careful removal of
solution from agarose resin after pelleted by centrifugation and b) eluting IP-complex by boiling the resin in denaturing
sample loading buffer for SDS-PAGE analysis, which will denature and separate antibody into heavy and light subunits.
Column method: Solution can be easily separated from the beads during the wash steps and the antibody-antigen complex
can be eluted by low pH buffer and neutralized for subsequent analysis instead of being denatured or cleaved into separate
subunits. Furthermore, the resin often can be reused to save the cost.
Highlights:
Efficiently and effectively remove contaminating proteins from beads
No resin loss, high recovery of antigen and co-precipitated proteins
Low pH elution provides milder and less denaturing recovery of the antibody-antigen complex
Fast and Easy with Spin protocol
Cost efficient by reusing affinity resin
Filter Disc does not bind to DNA/RNA or Proteins
Applications:
Immunoprecipitation (IP) Co-immunoprecipitation (co-IP) Immunodepletion
Other small scale affinity purification (affinity chromatography) such as Ni-resin, GST resin etc.