INSTRUCTIONS
Instructions for product no
A0-PU1-020
2000U
Version 15.1.2
Cleavage of up to 2.0mg IgG
Product Description
FabULOUS® (SpeB) is an enzyme for digestion of IgG in the upper hinge region. All subclasses of human (except hIgG4),
mouse, rat, goat and sheep IgG are digested by FabULOUS® yielding Fab and Fc fragments. The primary digestion site on
human IgG1 is ..KTHT / CPPCPAPE.. and on mouse IgG1 ..KPCIC / TVPES..
Incubation for one hour at 37°C under reducing conditions is generally applicable. FabULOUS® digests IgG in commonly used
buffers with pH ranging from 6.5 to 8.0 (Table 1). Best activity is obtained at 37°C.
FabULOUS® requires reducing agent in the digestion reaction (Table2). As reducing agent is present during reaction it is likely
that interchain thiols will be reduced.
The molecular weight of FabULOUS® is 28 724kDa and the enzyme contains a His-tag.
Content and storage
FabULOUS® is formulated in 10mM Tris, 150mM NaCl, pH 7.4, without preservatives, and supplied as lyophilized powder.
FabULOUS® is shipped on ice. Vials should be stored at -20°C upon arrival.
After reconstitution FabULOUS® is stable for 1 month at +4-8°C.
FabULOUS® is for R&D use only.
Unit Definition
1 unit FabULOUS® cleaves 1µg IgG when used as described in section below.
Additional Materials Required
•
•
•
Digestion Buffer: see Table 1
Reducing agent: see Table 2
Test tubes
Table 1. Buffers tested for compatibility with FabULOUS® digestion at different pH.
Compatible buffers
pH range
Phosphate buffered saline (PBS)
6.5 – 8.0
Tris buffer
7.0 – 8.0
Sodium acetate buffer
6.5 *
*below pH 6.5 increase the amount of enzyme
Table 2. Reducing agents tested with FabULOUS®.
Reducing agent
Concentration (mM)
pH
DTT
TCEP
1-5
1-5
6.5 - 8.0
Cysteine
Cysteamine
Mercaptoethanol
50-100
50-100
50-100
6.5 - 8.0
6.5 - 8.0
6.5 - 8.0
6.5 - 8.0
Customer
Customer Service
Service USA
USA Phone:
Phone: 617-444-8421
617-444-8421 || [email protected]
[email protected] || Genovis
Genovis AB,
AB, 245
245 First
First Street,
Street, Cambridge,
Cambridge, MA
MA 02142,
02142, USA
USA
Customer Service Europe Phone: 0046 (0)46 10 12 30 | [email protected] | Genovis AB, Box 790, SE-220 07 Lund, Sweden
Support [email protected] Order [email protected] | Fax: 0046 (0)46 12 80 20
www.genovis.com
Method
Cleavage of IgG
•
Make sure your antibody is in cleavage buffer (Table 1) with a concentration of 0.5-10 mg antibody/ml.
•
Reconstitute FabULOUS® in 50µl double distilled H2O, this leaves a concentration of 40 Units/µl.
•
Add FabULOUS® to the IgG. Add 1 Unit FabULOUS® /1µg IgG.
•
Add reducing agent to the digestion reaction, 1-5mM DTT or 1-5mM TCEP at neutral pH is normally used. See Table 2.
for different reducing agents.
•
Incubate for 1h at 37°C.
Quality Control
FabULOUS® is tested to ensure lot-to-lot consistency.
FabULOUS® is tested for absence of microbial contamination with blood agar plates, Sabouraud dextrose agar plates and fluid
thioglycollate medium.
®
FabULOUS
©2015 Genovis AB. All rights reserved. Aspects of FabULOUS® technology are encompassed by pending patent applications in
the name of Genovis AB.
The trademark FabULOUS is the property of Genovis AB.
For research use only. Not intended for any animal or human therapeutic diagnostic use.
Customer
Customer Service
Service USA
USA Phone:
Phone: 617-444-8421
617-444-8421 || [email protected]
[email protected] || Genovis
Genovis AB,
AB, 245
245 First
First Street,
Street, Cambridge,
Cambridge, MA
MA 02142,
02142, USA
USA
Customer Service Europe Phone: 0046 (0)46 10 12 30 | [email protected] | Genovis AB, Box 790, SE-220 07 Lund, Sweden
Support [email protected] Order [email protected] | Fax: 0046 (0)46 12 80 20
www.genovis.com