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Thermo Scientific™ Pierce™ F(ab')2 Micro Preparation Kit
Description
The Thermo Scientific Pierce F(ab')2 Micro Preparation Kit uses immobilized pepsin protease to selectively digest whole human and other IgG antibodies to make F(ab')2 fragments that retain antigen binding activity.
Features of the F(ab')2 Micro Preparation Kit:
- Ideal when sample size is limited—Use the micro kit for 25 to 250 μg IgG
- Enzyme-free digestion products—Immobilized Pepsin (beaded agarose resin) provides for control of the digestion reaction and complete removal of resulting antibody fragments from the proteolytic enzyme
- Suitable for human and other species of IgG—the kit procedure is optimized for human, rabbit and mouse IgG. Pepsin-based digestion is effective for other species and subclasses of IgG, although the purification step requires that the antibody effectively binds to Protein A. (Note: for best results with mouse IgG1, use Part No. 44980.)
- Complete—kits include all reagents needed to prepare and purify antibody fragments
- Fast—spin format greatly reduces sample processing time
- Flexible—protocols are included for multiple species and IgG subclasses, as well as sample size and concentration
- Efficient—enhanced yield and sample purity
These F(ab')2 Preparation Kits are suitable for human, rabbit, mouse and other species and subclasses of IgG except mouse IgG1 (Cat. No. 44680). The antibody digestion reaction with pepsin agarose is performed in convenient disposable spin columns that allow efficient removal of the immobilized protease and maximum recovery of the IgG fragments. Also included in the kits are Thermo Scientific NAb Protein A Spin Columns and buffers to efficiently purify the resulting fragments. Protein A binds the Fc fragments and undigested IgG, allowing the pure F(ab')2 fragments to be recovered in the flow-through fraction. The kits also include Thermo Scientific Zeba Desalting Spin Columns for preparing the IgG sample quickly without dilution instead of utilizing time-consuming dialysis steps.
The kits use pepsin, a nonspecific endopeptidase, to enzymatically digest the Fc portion of whole IgG to yield the fragment known as F(ab')2. This fragment is composed of a pair of Fab' units connected by two disulfide bonds. Because the pepsin protease is supplied in immobilized form as beaded agarose resin, the digestion reaction is easily stopped by removing the resin from the IgG solution; the result is digest products that are enzyme-free.
Related Products
- Pierce™ F(ab')2 Preparation Kit (Cat. No. 44988)
- Pierce™ Fab Micro Preparation Kit (Cat. No. 44685)
- Pierce™ Mouse IgG1 Fab and F(ab')2 Micro Preparation Kit (Cat. No. 44680)
Specifications
Specifications
| Product Line | Pierce |
| Quantity | 10 Antibody Samples, each containing 25 to 250 μg IgG |
| Kit Contents | Pierce F(ab')2 Micro Preparation Kit |
| Content And Storage | Sufficient For: 10 antibody samples, each containing 25 to 250 μg IgG • Pierce Pepsin Agarose, 0.5 mL • F(ab')2 Digestion Buffer, 55 mL • NAb Protein A Plus Spin Columns, 0.2 mL, 2 columns • PBS Packs (each makes 500 mL), 2 packs • 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°C |
| Target Gene | ERK5/BMK1 (pan) |
Frequently Asked Questions (FAQs)
No. For best results, use the immobilized enzyme once and discard after use.
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.
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.)
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
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
For Research Use Only. Not for use in diagnostic procedures.