Pierce™ Antibody Clean-up Kit - FAQs

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3 product FAQs found

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.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.