Melon™ Gel Chromatography Cartridges, 1 mL, 2 x 1 mL - FAQs

For serum, a volume double the Melon Gel Support volume is the maximum amount that can be processed at one time. To determine if too much sample was processed, examine the purified IgG by SDS-PAGE for additional protein bands.

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No. Thermo Scientific Thiophilic Adsorbent (Cat. Nos. 20500 and 44916) can be used to purify IgY from serum. The Chicken IgY Purification Kit (Cat. No. 44918) can be used to purify IgY from egg yolks.

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Yes. With the Melon Gel IgG Spin Purification Kit, 90% recovery of IgG with greater than 80% purity are typically observed for a single purification. Although IgA and IgM may be present along with the purified IgG, a high percentage (> 80%) of IgA and IgM is retained on the gel support. This high binding of IgA and IgM to the gel support combined with the fact that IgA and IgM are present in low abundance in serum results in low levels of these antibody molecules in the purified IgG. Here are other advantages of using the Melon Gel IgG Spin Purification Kit over traditional Protein A or Protein G methods:
-Simple, one-step protocol - IgG purification with Melon Gel Resin does not involve binding, washing, and multiple elution steps required by Protein A and Protein G
-Rapid purification - antibody purification from serum is four to six times faster than Protein A or G methods
-Minimal dilution - reduced need to concentrate antibody before storage or use
-Robust purification - Melon Gel IgG Purification Kits work with a wide range of antibodies including many that do not purify well on Protein A or Protein G
-Gentle purification - No harsh antibody elution conditions helps retain IgG activity
-Regenerable support - Melon Gel Resin can be used for multiple antibody purifications

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The Melon Gel Support can be used up to three times. Performing an ammonium sulfate precipitation on serum samples containing large amounts of non-antibody protein before purification improves purification efficiency and increases reusability.

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No. However, researchers may want to add an equal volume of binding buffer to elevate the pH towards neutral.

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Yes. Dialyze or desalt the antibody solution against the Purification Buffer provided in the kit. The undiluted antibody solution can then be added to the Melon Gel Support using the volumes indicated in the instructions. Tech Tip # 55 (https://assets.thermofisher.com/TFS-Assets/LSG/Application-Notes/TR0055-Melon-Gel-for-BSA.pdf) provides a protocol for this method. Another option is to use Cat. No. 44600 Thermo Scientific Antibody Clean-Up Kit for removing BSA, gelatin and other stabilizer proteins from antibody stock solutions. Sufficient for purifying 10 x 0.5 mL IgG samples.

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Microcentrifuge spin cups are provided in the Melon Gel IgG Spin Purification Kit (Cat. No. 45206). No columns are included in the other Melon Gel Kits (Cat. No.s 45212 and 45214) so that researchers can adjust the scale of their purification. A wide variety of disposable columns are available for use with these kits. The product instructions for the Monoclonal Kit include a procedure for using a filter funnel and flask instead of a column for large-scale (e.g., 1 liter purifications.

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Yes, ascites samples must be treated with the Ascites Conditioning Reagent (Cat. No. 45219) before antibody purification. See the instructions and other product information for the Monoclonal IgG Purification Kit (Cat. No. 45214).

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There are three different formats. The Melon Gel IgG Purification Kit (Cat. No. 45212) and Melon Gel IgG Spin Purification Kit (Cat. No. 45206) are for purifying antibodies from serum. The Melon Gel Monoclonal IgG Purification Kit (Cat. No. 45214) is for purifying monoclonal antibodies from cell-culture supernatant and ascites fluid. Ready-to-use prepacked chromatography cartridges are available as two 1 mL columns (Cat. No. 89932) or one 5 mL column (Cat. No. 89933).

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At physiological pH, Melon Gel Support binds non-antibody proteins in serum, such as albumin and transferrin, allowing the antibody to flow through in a mild buffer suitable for storage and downstream applications.

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The Melon Gel IgG Spin Purification Kit is designed for the purification of IgG from serum samples. 90% recovery of IgG with greater than 80% purity are typically observed for a single purification. Although IgA and IgM may be present along with the purified IgG, a high percentage (> 80%) of IgA and IgM is retained on the gel support. This high binding of IgA and IgM to the gel support combined with the fact that IgA and IgM are present in low abundance in serum results in low levels of these antibody molecules in the purified IgG.

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

For purification of antibodies, we offer Melon Gel, which binds other proteins from serum, ascites or culture supernatant, allowing the IgG to be collected in the flow-through. As low pH is typically used to elute IgG from Protein A, G, A/G or L, the use of Melon Gel can spare the IgG from exposure to such harsh conditions, which can be particularly important for monoclonal antibodies.

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