Melon Gel Monoclonal IgG Purification Kit
It’s almost as easy as having someone
else do the work for you!
The new Melon™ Gel Monoclonal IgG Purification Kit from Pierce offers a fast and simple
protocol for purifying IgG from cell culture supernatants and ascites that renders Protein A
and Protein G methods obsolete
How does Melon™ Gel work?
Melon™ Gel is composed of a proprietary combination of ligands that
retains the component proteins typically found in serum and allows
IgG to pass through.
The Melon™ Gel Monoclonal IgG Purification Kit:
• Replaces tedious bind-and-release antibody-isolation methods,
resulting in a fast, easy and gentle approach to antibody recovery
and purification
• Delivers monoclonal antibodies (MAbs) with quality and purity
equal to – or better than – that obtained using classical Protein A
or Protein G purification methods
• Purifies monoclonal IgG from up to 1 L of cell culture supernatant
(serum-containing or serum-free) and up to 200 ml of ascites fluid
Figure 1. Melon™ Gel offers better recovery and purity from cell culture super-
natant than Protein G. IgG was purified from cell culture supernatant containing
10% fetal bovine serum (FBS), resolved by SDS-PAGE and detected with Imperial™
Protein Stain† (Product # 24615). Lane 1: Original cell culture supernatant, Lane 2:
IgG after precipitation with Saturated Ammonium Sulfate (Product # 45216), Lane
3: Melon™ Gel-purified IgG and Lanes 4-6: Protein G-purified IgG.
www.piercenet.com/melg92h
Highlights:
• Fast – purify antibodies in half the time of Protein A or Protein G
• Efficient – recovery and purity is often better than Protein A or
Protein G purification
• Robust – effective for antibodies known not to bind to Protein A or
Protein G
• Gentle – antibody retains activity because it is not exposed to
denaturing conditions
• Regenerable – support can be used for up to six purifications
• Compatible – buffer is amine-free and does not require removal
or neutralization
• Flexible – accommodates a broad range of sample volumes
Cell culture: Spin columns can process supernatant samples that
are < 30 ml. Filter flasks allow processing of samples up to 1 L.
Ascites: Spin columns process samples up to 6 ml.
Figure 2. Melon™ Gel offers better recovery and purity from ascites fluid than
Protein G. IgG1 was purified from ascites fluid, resolved by SDS-PAGE and detect-
ed with GelCode™ Blue Stain Reagent (Product # 24590). Lane 1: Original ascites
fluid, Lane 2: treated with Ascites Conditioning Reagent (Product # 45219) and
purified with Melon™ Gel, Lane 3: Protein G column flow-through, Lanes 4-5:
Protein G column washes and Lanes 6-8: Protein G column elutions. NOTE: The
Protein G-purified sample is contaminated with transferrin (Lanes 6-8). The
Melon™ Gel-purified sample is not (Lane 2).
 Cell Culture Supernatant Purification Ascites Purification
Slide-A-Lyzer® 2
1. Combine the Ascites 2. Centrifuge sample at
1

Cassette

Conditioning Reagent with 5,000 x g for 10 minutes.

1. Buffer exchange 2 x 1 hour. purification buffer and
4
then add to the sample.
3

U.S. Pat.
U.S. # 5,503,741
Pat. # 5,503,741

Mix end-over-end for

10 minutes.

2. In a funnel, drain storage buffer from resin and

then transfer funnel to a new filter flask. Add 3. Remove Ascites Conditioning
buffer-exchanged sample and stir for 5 minutes. Slide-A-Lyzer® 2
Reagent via dialysis or desalting.

1
Cassette

3
U.S. Pat.
U.S. # 5,503,741
Pat. # 5,503,741

4. In a spin column, combine 5. Centrifuge to collect

buffer-exchanged sample purified sample.
3. Apply vacuum to collect the purified sample in with gel and incubate
the new flask. end-over-end for 5 minutes.

0.45 Ordering Information

0.40 PKG.
PRODUCT # DESCRIPTION SIZE
0.35
Absorbance at 450 nm

45214 Melon™ Gel Monoclonal IgG Purification Kit† Kit

0.30 Sufficient reagents to purify IgG from up to 1 L of cell
0.25 culture supernatant or up to 200 ml of ascites fluid.
Includes: Melon™ Gel IgG Purification Support, 200 ml settled gel
0.20 Melon™ Gel Purification Buffer, 100X, dry mix
0.15 Melon™ Gel Regenerant, 5X, dry mix
0.10 Melon™ Gel 45216 Saturated Ammonium Sulfate Solution 1L
0.05 Protein G
For use with the Melon™ Gel Monoclonal IgG
Purification Kit and Melon™ Gel IgG Purification Kits
0.00 for Sera or as a general-purpose salting-out agent
0.00 1.00 2.00 3.00 4.00 for protein-preparation applications.
Amount of KLH (µg/well) 45219 Ascites Conditioning Reagent 5 ml
For use with the Melon™ Gel Monoclonal IgG
Figure 3. Melon™ Gel provides greater IgG recovery than Protein G. Anti- Purification Kit.
KLH IgG3 produced in cell culture was purified using Melon™ Gel and Protein G Instructions not included. Use of this reagent is described in the
agarose. Recoveries were determined by applying the Melon™ Gel- and Protein instructions that are provided with Product # 45214.
G-purified antibodies to KLH-coated plates and detecting with HRP-labeled goat †
U.S. patent pending on Melon™ Gel Monoclonal IgG Purification Technology and
anti-mouse antibody (Product # 31430) and 1-Step™ Ultra TMB-ELISA (Product Imperial™ Protein Stain Technology.
# 34028). The absorbances of the purified samples were compared. Recovery of
antibodies produced in serum-free media and ascites fluid were also greater
than or equal to recoveries obtained with Protein G.

Tel: 815-968-0747 or 800-874-3723 • Fax: 815-968-7316 • Customer Assistance E-mail: CS@piercenet.com • www.piercenet.com # 1601296 09/05
© Pierce Biotechnology, Inc., 2005. Pierce products are supplied for research or manufacturing applications only.