Mouse IgG1 Fab and F(ab′)2 Preparation Kit
| Cat No: | BIOK2025 |
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| Source : | N/A |
| Application : | Fragmentation |
| Shipping : | The product is shipped at room temperature. |
| Storage : | Store at 2-8℃. |
| Note : | For research use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. |
| Background : | Mouse IgG1 Fab and F(ab′)2 Preparation Kit uses Immobilized Ficin to prepare fragments from mouse IgG1. Ficin generates F(ab′)2 fragments exclusively in the presence of 1-4mM cysteine; and Fab fragments are generated in the presence of 25mM cysteine. Fragment generation from other IgG species and isotypes might be possible by modifying the cysteine concentration and other digestion parameters. Pepsin is commonly used for generating F(ab′)2 fragments because the pepsin cleavage site on human IgG contains a Leu 234, which is conserved in most species; however mouse IgG1 lacks this residue and others, which possibly contributes to the restricted hinge region and resistance to pepsin cleaveage. Also, the low pH required for pepsin digestion can destroy or damage antibodies. For comparison, mouse IgG1 monoclonal antibodies were digested with ficin, pepsin, bromelain and elastase. Ficin digestion produced high yields of F(ab′)2 fragments with the highest residual antigen-binding activity and immunoreactivity. Affinity constants of ficin-generated F(ab′)2 fragments were near those of intact antibody. This kit contains the necessary components for Fab or F(ab′)2 generation of mouse IgG1 and subsequent purification. Immobilized Ficin enables immediate cessation of the digestion by simply removing the resin from the antibody digest solution. The included Spin Columns allow easy manipulation of the resin and maximum Fab and F(ab′)2 recoveries. The prepacked immobilized Protein A spin column and optimized binding buffer binds the intact Fc fragments and undigested IgG, allowing for efficient Fab or F(ab′)2 fragments purification. The optimized cysteine concentration produces Fab or F(ab′)2 with maximum purity. This complete kit makes Fab and F(ab′)2 generation and purification simple, fast and effective. |
| Keywords : | Fab |
| Synonyms : | Mouse IgG1 Fab and F(ab')2 Micro Preparation Kit; 小鼠 IgG1 Fab 和 F(ab') 2 微型制备试剂盒; 小鼠 IgG1 Fab & F(ab') 2 制备; Fragmentation; F(ab)2 制备试剂盒; Fab 制备试剂盒; Fab2 制备试剂盒; 抗体片段试剂盒 |
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