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Description
Spe BProduct Specification Species Streptococcus pyogenes Synonyms Streptopain, Exotoxin type B, Group A streptococcal cysteine protease (Streptococcal cysteine proteinase), SPE B, Streptococcus peptidase A (SPP) Expression System E. coli Molecular Weight 29kDa (Reducing) Purity 95% by SDS PAGE&HPLC Tag His Tag Physical Appearance Liquid Storage Buffer 20mM Tris HCl, 0. 5M NaCl, 50% Glycerol (pH 7. 0) Stability & Storage Store at 25 ~ 15 for 2 years
Product Specification
| Species | Streptococcus pyogenes |
| Synonyms | Streptopain, Exotoxin type B, Group A streptococcal cysteine protease (Streptococcal cysteine proteinase), SPE B, Streptococcus peptidase A (SPP) |
| Expression System | E.coli |
| Molecular Weight | 29kDa (Reducing) |
| Purity | ＞95% by SDS-PAGE&HPLC |
| Tag | His Tag |
| Physical Appearance | Liquid |
| Storage Buffer | 20mM Tris-HCl, 0.5M NaCl, 50% Glycerol (pH 7.0) |
| Stability & Storage | Store at -25 ~ -15℃ for 2 years |
| Reference | [1] Musser J M , Stockbauer K , Kapur V ,et al.Substitution of cysteine 192 in a highly conserved Streptococcus pyogenes extracellular cysteine protease (interleukin 1beta convertase) alters proteolytic activity and ablates zymogen processing.[J].Infection & Immunity, 1996, 64(6):1913. |
Background
SpeB is a unique recombinant cysteine protease that can cleave the hinge regions of various species and subclasses of antibodies, thereby generating a homogeneous pool of Fab fragments. This enzyme requires reducing conditions when acting on IgG. SpeB can digest human, mouse, rat, goat and sheep IgG, thereby producing Fab and Fc fragments. The main digestion site of human IgG1 is KTHT/CPPCPAPE.
Components
UA070131: 40U/µL SpeB in 20mM Tris-HCl, 0.5M NaCl, 50% Glycerol (pH 7.0)
5*Reaction Buffer: 5*PBS, 25mM DTT (pH 7.0)
Protocol
1.Take up to 10µg of human IgG1, adjust to 39µl with purified water;
2.Add 10µl of 5*Reaction Buffer;
3.Add 1µl SpeB to a total volume of 50µl, mix gently;
4. React for 1 hour at 37°C.
Guidelines
1. For different antibodies sample, the optimal enzyme concentration and reaction time need to be mapped experimentally;
2. Higher enzyme concentrations increase the digestion efficiency of individual antibodies and need to be optimized accordingly.
Unit Definition
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