Catalog No.
YXX05101
Description
IdeZ Protease is a recombinant antibody specific protease that recognizes all human, sheep, monkey, and rabbit IgG subclasses, cleaving specifically at a single recognition site below the hinge region, to yield a homogenous pool of F(ab´)2 and Fc fragments. IdeZ Protease more effectively cleaves murine IgG2a and IgG3 than IdeS.
Biological activity
One unit will cleave ≥95% of 1μg of recombinant monoclonal IgG in 30 minutes at 37°C.
Expression system
E. coli
Species
Streptococcus equi
Protein length
IdeZ Protease is cloned from Streptococcus equi and expressed in E.coli.
Predicted molecular weight
37.75 kDa
Nature
Recombinant
Target
IdeZ Protease (IgG Specific)
Concentration
50000U/ml
Endotoxin level
Please contact with the lab for this information.
Purity
>95% as determined by SDS-PAGE.
Applications
Proteomics
Form
Liquid
Storage buffer
0.01M PBS, pH 7.4, 50% glycerol
Experimental Procedure
Protocol
1. Add the desired amount of IgG (up to 5mg) in digestion buffer or other compatible buffer.
2. Add IdeZ Protease to the IgG sample: Add 1 unit of IdeZ Protease per 1μg of IgG to be digested.
For example, add 1μl (50 units) of IdeZ Protease to digest 50μg of IgG.
3. Incubate sample at 37°C for 30–60 minutes.
IdeZ Protease is most active in buffers at or near neutral pH. The recommended digestion buffer is 50mM sodium phosphate, 150mM NaCl (pH 6.6), but many common biological buffers such as Tris or PBS also can be used. Buffers outside this pH range (e.g., acetate buffers) may be used, but the incubation time and/or enzyme amount may have to be optimized on a case-by-case basis.
Mouse IgG2a and IgG3 typically require 2–4 hours for complete digestion. In some cases, human IgG2 may require incubation beyond 60 minutes.
Notes
IgG concentration ideally should be in the range of 1–20mg/ml.
IdeZ Protease efficiently cleaves human, humanized, chimeric, sheep, rabbit and monkey IgGs as well as mouse IgG2a and IgG3.
IdeZ Protease also will cleave many Fc-fusion proteins as well as antibody-drug conjugates (ADCs).
IdeZ Protease does not cleave mouse IgG1/IgG2b, rat, porcine, bovine or goat IgG. IdeZ Protease also does not cleave non-IgG isotypes including IgA, IgM, IgD and IgE.
Shipping
In general, proteins are provided as lyophilized powder/frozen liquid. They are shipped out with dry ice/blue ice unless customers require otherwise.
Stability and Storage
Use a manual defrost freezer and avoid repeated freeze thaw cycles.Store at 2 to 8 °C for one week .Store at -20 to -80 °C for twelve months from the date of receipt.
For research use only. Not for human or drug use.
