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Thermo Scientific™ Pierce™ Antibody Clean-up Kit

Catalog No. PI44600
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PI44600 10 Reactions
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Catalog No. PI44600 Supplier Thermo Scientific™ Supplier No. 44600
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Description

Remove BSA, gelatin and other stabilizer proteins from antibody stock solutions before labeling and conjugation procedures.

Thermo Scientific™ Pierce Antibody Clean-Up Kit is for removing BSA, gelatin and other stabilizer proteins from antibody stock solutions before 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). 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.

Highlights:

  • 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

Includes:

Melon Gel IgG Purification Support and Buffer, desalting columns, microcentrifuge spin columns and tubes

Specifications

Product Type Antibody Clean-up Kit
Product Line Pierce
Column Type Proprietary Resin, Affinity
Format Kit, Microcentrifuge Spin Column
Stationary Phase Proprietary Ligand
Purification Target IgG Antibodies
Content And Storage Sufficient For: Purifying 10 x 0.5 mL IgG samples• Melon Gel IgG Purification Support, 1.2 mL
• Melon Gel Purification Buffer, 100 mL
• Zeba Spin Desalting Columns, 7K MWCO, 2 mL, 10 columns
• Spin Columns, 0.8 mL, 10 columns
• Microcentrifuge Tubes, 2 mL, 30 tubes

Store at 4°C
Quantity 10 Reactions

Frequently Asked Questions (FAQs)

Will glycerol pose a problem when purifying antibodies with the Pierce Antibody Clean-up Kit (Cat. No. 44600)?

When using the Pierce Antibody Clean-up Kit, glycerol may pose a problem for the spin desalting columns for buffer exchange, but not for the Melon Gel itself. If glycerol concentrations are high, the solution becomes viscous enough to exert a frictional drag during spin column chromatography and the performance and yield may suffer. Please find a technical tip on using Melon Gel for cleaning up BSA or gelatin from commercial antibodies.

If the glycerol concentration is higher than 10%, dialysis may be considered to exchange the sample into the Melon Gel buffer. We have some dialysis devices suitable for small volume dialysis, e.g., the Slide-A-Lyzer MINI devices: see here. Alternatively, some of the smaller Slide-A-Lyzer dialysis cassettes can be used: see here.
I want to use the Antibody Clean-Up Kit (Cat. No. 44600) with a purified antibody that has a "protein based stabilizer". Do I need to first perform the Zeba desalting step A in the kit, or can I proceed directly to the BSA/Gelatin Removal step B?

The Zeba desalting step A is necessary as it results in a buffer exchange into the Melon Gel Purification Buffer. Having the antibody in this buffer is critical for the functionality of the Melon Gel used to remove protein.
I typically use BSA or gelatin to store my purified antibody solution for long-term storage. I'm worried that it will interfere with my immunodetection experiment. Is there a way to remove BSA or gelatin from my antibody solution?

The Pierce Antibody Clean-up Kit (Cat. No. 44600) was specifically designed for removal of amine containing molecules from IgG. Please view this Tech Tip (https://tools.thermofisher.com/content/sfs/brochures/TR0055-Melon-Gel-for-BSA.pdf) for additional strategies on how to remove BSA and gelatin from antibody solutions.

For Research Use Only. Not for use in diagnostic procedures.