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Thermo Scientific™ Pierce™ Mouse IgG1 Fab and F(ab')2 Micro Preparation Kit

Catalog No. PI44680
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10 Antibody Samples
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Catalog No. Quantity
PI44680 10 Antibody Samples
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Catalog No. PI44680 Supplier Thermo Scientific™ Supplier No. 44680
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Description

Efficiently digest mouse IgG1 antibodies into Fab or F(ab')2 fragments with immobilized ficin protease.

The Thermo Scientific Pierce Mouse IgG1 Fab and F(ab')2 Micro Preparation Kit uses immobilized ficin protease to efficiently digest mouse IgG1 into Fab or F(ab')2 fragments, depending on the chosen reaction conditions.

Features of the Mouse IgG1 Fab and F(ab')2 Micro Preparation Kit:

  • Capacity—kit provides for fragmenting 25 to 250 μg IgG
  • Complete—kit includes all reagents needed to prepare and purify antibody fragments
  • Enzyme-free digestion products—Immobilized Ficin (beaded agarose resin) provides for control of the digestion reaction and complete removal of resulting antibody fragments from the proteolytic enzyme
  • Optimized for mouse IgG1—the kit procedure is optimized for mouse IgG1; the only effective method for this IgG species and subclass

Related Products

  • Pierce™ Mouse IgG1 Fab and F(ab')2 Preparation Kit (Cat. No. 44980)
  • Pierce™ F(ab')2 Micro Preparation Kit (Cat. No. 44688)
  • Pierce™ Fab Micro Preparation Kit (Cat. No. 44685)

Specifications

Product Line Pierce
Quantity 10 Antibody Samples
Kit Contents Pierce Mouse IgG1 Fab and F(ab')2 Micro Preparation Kit
Content And Storage Sufficient For: 10 antibody samples, each containing 25 to 250 μg IgG
• Pierce Ficin Agarose, 0.8 mL
• Cysteine-HCl, 1 g
• Mouse IgG1 Digestion Buffer, 120 mL
• NAb Protein A Plus Spin Columns, 0.2 mL, 2 columns
• Protein A Binding Buffer, 50 mL
• IgG Elution Buffer, 50 mL
• Zeba Spin Desalting Columns, 7K MWCO, 0.5 mL, 10 columns
• Spin Columns, 0.8 mL, 10 columns
• Microcentrifuge tubes, 2 mL, 30 tubes

Store at 4–8°C.

Frequently Asked Questions (FAQs)

Can I reuse the immobilized digestion enzymes in the Thermo Scientific IgG Fragmentation kit?

No. For best results, use the immobilized enzyme once and discard after use.
How long does IgG digestion take using the Thermo Scientific IgG Fragmentation kit?

The digestion time is a function of the IgG concentration and source. For rabbit, mouse and human IgG, optimized digestion times are given in the instructions for each kit. Digestion times listed in the product instructions result in 80-90% digestion, although efficiency varies depending on IgG preparation and source.
In the Thermo Scientific IgG Fragmentation kit, when digesting an IgG that does not bind to Protein A, how do I purify the Fab and F(abÁ)2 fragments?

If the IgG is a species or subclass that does not bind to Protein A, researchers can immobilize an antibody specific for the desired fragment. For example, to purify digested rat IgG2a, use the Aminolink Plus Immobilization Kit (Cat. No. 44894) to immobilize goat anti-rat Fc antibody. Our Tech Tip #34 and other web resources list IgG binding properties of Protein A. (Because Protein G and Protein A/G bind Fab fragments, they are not effective for this application.)
Can I purchase the immobilized enzymes (proteases) from the Thermo Scientific IgG Fragmentation kit separately?

Yes. The three immobilized enzymes are available in 5 mL package sizes as shown below:

• Immobilized Ficin, Cat. No. 44881
• Immobilized Papain, Cat. No. 20341
• Immobilized Pepsin, Cat. No. 20343
How many Thermo Scientific IgG Fragmentation kits are available and which enzymes are included?

There are six Thermo Scientific IgG fragmentation kits:

• Fab Preparation Kit (Papain), Cat. No. 44985
• Fab Micro Preparation Kit (Papain), Cat. No. 44685
• F(abÁ)2 Preparation Kit (Pepsin), Cat. No. 44988
• F(abÁ)2 Micro Preparation Kit, (Pepsin), Cat. No. 44688
• Mouse IgG1 Fab and F(abÁ)2 Preparation Kit (Ficin), Cat. No. 44980
• Mouse IgG1 Fab and F(abÁ)2 Micro Preparation Kit (Ficin), Cat. No. 44680
What species of IgG can be fragmented with the Thermo Scientific IgG Fragmentation kits?

The protocols are optimized for rabbit, human and mouse IgG. Fragmentation of other IgG species may require optimization. To purify Fab or F(abÁ)2 fragments from the digest, the IgG species must be able to bind to Protein A. For mouse IgG1, use a ficin-based kit (Cat. Nos. 44980, 44680).

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