The Thermo Scientific Pierce Antibody Clean-Up Kit is for removing BSA, gelatin and other stabilizer proteins from antibody stock solutions before amine-based chemical labeling and conjugation procedures.
This is a complete and easy-to-use kit for removing BSA and gelatin stabilizing proteins (up to 1%) from IgG samples. The protocol uses Thermo Scientific Melon Gel to bind these frequently used storage proteins, allowing the antibody to flow through in a purified form. The antibody is recovered in an amine-free and carboxyl-free buffer, ready for use in downstream applications such as biotinylation, fluorescent labeling, crosslinking, immobilization and enzymatic fragmentation (e.g., Fab preparation).
- Fast – removes carrier protein from antibody solutions in half the time required for purifying antibodies through Protein A or G purification procedures
- Gentle – no need to neutralize or desalt to mitigate the effects of harsh elution buffers
- Complete – supplied desalting columns provide for easy buffer exchange into the required purification buffer or quick exchange into final buffer of choice
- Economical – sufficient reagents to perform 10 antibody clean-up reactions
Bovine serum albumin (BSA) and gelatin are often added to purified antibody solutions as stabilizers for long-term storage. Inclusion of 0.2 to 1% (2 to 10mg/mL) BSA or gelatin helps stabilize dilute antibody solutions. Generally these added proteins do not interfere with immunodetection methods; however, their presence in solution does interfere with amine-reactive conjugation chemistries that are the basis for most biotinylation, fluorescent dye labeling, antibody fragmentation and covalent immobilization methods. For such methods, the antibody must be isolated from the stabilizing proteins, and the Antibody Clean-up Kit accomplishes this efficiently.
Efficient removal of bovine serum albumin (BSA) from mouse IgG samples. Mouse IgG1 (0.25mg/mL) samples were processed using The Thermo Scientific Pierce Antibody Clean-up Kit. Samples (10µL each) were subjected to SDS-PAGE and stained with Imperial Protein Stain.
Lane 1: MW marker
Lane 2: Pure IgG without BSA
Lanes 3-5: IgG samples containing 0.75%, 0.5% and 0.25% BSA, respectively
Lanes 6-8: Same IgG samples as lanes 3-5 after after performing the kit procedure.
Review of antibody purification methods
Biotin-labeling Reagents and Kits
Fluorescent Labeling Reagents and Kits
Immunoprecipitation Reagents and Kits
Supports for Covalent Protein Immobilization
Melon Gel Antibody Purification Kits