Plant organelle/membrane isolation
- Mitochondrial fraction
- Nuclear fraction
- Plasma membrane fraction
- Thylakoid extraction
- Arabidopsis thylakoid preparation
- Arabidopsis thylakoid extraction
- Arabidopsis lumen extraction
- BBY preparation
- Chlorophyll measurements
- PSII RC extraction for cryo-EM
- Extraction of leaf proteins
- Diatom protein extraction
- Phenol protein extraction
- Protein extraction from grasses
- Ponceau membrane staining
- TCA acetone precipitation method
- Western blot protocol
- Western blot video tutorial
- Peptide neutralization/competition assay
- Simultaneous Western blot
- Quantitative Western blot
- Quantitative Western blot video tutorial
- Western blot troubleshooting
- Western blot using IgY
- Dot blot
- ELISA
- Immunoprecipitation
- Immunoprecipitation/IgY
- Immunohistochemistry
- Anti-KLH antibody removal
- Yolk delipidation
Technical information
Antibody typesPurification
- Antibody purification
- Antibody purification - small amount of protein
- IgY purification methods
- Protein purification using antibodies
- Elution of antibodies from affinity columns
Technical information > Elution of antibodies from affinity columnsLimitations
Elution of antibodies1. Antibodies are purified based on their affinity to Protein A or Protein G to obtain a total immunoglobulin pool. Binding to the matrix occurs mainly via Fc part of the antibody.
2. Either antigen or antibody is immobilized on the appropriate matrix. The bond between antibody and antigen needs to be disrupted to allow the elution.
General commentsElution conditions must be tested experimentally and established individually for each antigen-antibody pair. In some cases, effective elution cannot be achieved by standard conditions (low pH), and other methods must be employed. However, if using harsher methods, eluted proteins can be irreversibly denatured. Elution conditions are a compromise between the achieved yield and protein activity.Theoretical yields of affinity purifications
|