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Thermo Scientific™ Melon™ Gel Monoclonal IgG Purification Kit

Catalog No. PI45214
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PI45214 200 mL
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Catalog No. PI45214 Supplier Thermo Scientific™ Supplier No. 45214
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Description

Purify mouse monoclonal antibodies from serum, ascites or hybridoma cell culture supernatant with this high-yield, mild monoclonal IgG purification kit.

Thermo Scientific™ Melon Gel Monoclonal IgG Purification Kit is a high-yield, mild purification system for mouse monoclonal antibodies from serum, ascites or hybridoma cell culture supernatant.

The Melon Gel Monoclonal IgG Purification Kit consists of a large package size of Melon Gel Resin and an optimized protocol for easy and mild purification of monoclonal IgG antibodies from hybridoma culture supernatant or ascites fluid. Unlike typical bind-and-elute affinity methods, Melon Gel Purification is based on the binding of non-IgG proteins from a prepared supernatant or ascites sample, thereby allowing the pure IgG to be recovered in the flow-through (non-bound) fraction. Antibodies are recovered ready for use in assays or subsequent conjugation and other modification steps. There is no need to neutralize or desalt to mitigate the effects of harsh elution buffers. Melon Gel Purification saves time compared to classical Protein A or Protein G methods without compromising the quality of the antibody.

Highlights:

  • Fast – purify antibodies in half the time of required for Protein A or G

  • Efficient – recovery and purity is often better than Protein A or G purification

  • Robust – effective for subclasses of IgG that bind poorly to Protein A or G

  • Gentle – antibodies retain activity because they are not exposed to denaturing conditions

  • Reusable – support can be regenerated and used for up to 6 purifications

  • Compatible – amine-free buffer does not require removal or neutralization

  • Flexible – accommodates a broad range of sample volumes and processing vessels

Includes:

Melon Gel Resin, purification buffer and resin regenerant

Specifications

Product Type IgG Purification Kit
Product Line Melon
Column Type Proprietary Resin, Affinity
Format Kit, Bottle
Stationary Phase Proprietary Ligand
Purification Target IgG Antibodies
Description Melon Gel Monoclonal IgG Purification Kit
Content And Storage Sufficient For: IgG purification from up to 1 L of cell culture supernatant or up to 200 mL of ascites fluid
• Melon Gel Support, 200 mL
• Melon Gel Purification Buffer (dry mix to make 100X), 1 L
• Melon Gel Regenerant (dry mix to make 5X), 1 L

Store at 4°C
Quantity 200 mL

Frequently Asked Questions (FAQs)

How can I determine when the Melon Gel Support is saturated?

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.
Can Melon Gel purify chicken IgY?

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.
Is the Melon Gel IgG Spin Purification Kit (Cat. No. 45206) a viable alternative to using traditional Protein A and Protein G methods for purification of IgG from serum?

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
How many times can I reuse the Melon Gel Support?

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.
After Melon Gel purification, must I desalt or dialyze the purified antibody before using the Thermo Scientific Direct IP Kit (Cat. No. 26148)?

No. However, researchers may want to add an equal volume of binding buffer to elevate the pH towards neutral.
Can the Melon Gel Method remove BSA or gelatin from commercially available antibody?

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.

Are columns provided in the Melon Gel Purification kits?

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.
Does ascites fluid require any special treatment before processing using the Melon Gel Purification Kit?

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).
What formats are available for the Melon Gel Support?

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).
How does Melon Gel Purification work?

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.
Can I use the Melon Gel IgG Spin Purification Kit (Cat. No. 45206) to purify IgA and/or IgM from my serum samples?

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.
Do you offer a kit for antibody purification?

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.

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