HCA008 binds to the C-terminal 122 amino acids of mKIAA0051, also known as Murine IQ motif containing GTPase activating protein 1.
The mKIAA0051 protein (IQGAP1) has a fundamental role in cell motility and invasion. It binds to activated CDC42 but does not stimulate its GTPase activity. It also associates with calmodulin and regulates cadherin-based cell adhesion. Potentially mKIAA0051 serves as an assembly scaffold for the organization of a multimolecular complex that would interface incoming signals to the reorganization of the actin cytoskeleton at the plasma membrane.
HCA008 may bind to the human Ras GTPase-activating-like protein IQGAP1 (p195), due to 98% sequence identity in the antigen region.
Product Form
A lyophilised bivalent human recombinant Fab selected from the HuCAL®GOLD phage display library. Expressed inE. coliand purified using NiNTA affinity chromatography. This Fab fragment is dimerized via a helix-turn-helix motif. The antibody is tagged with a myc-tag (EQKLISEEDL) and a his-tag (HHHHHH) at the C-terminus of the antibody heavy chain.
Activity
Activity was tested by indirect ELISA: recombinant purified mKIAA0051 (5 µg/ml) plus unrelated control proteins were immobilized on a microtiter plate. Specific binding was monitored by first adding HCA008 (2 µg/ml), then adding a secondary antibody (goat anti-human F(ab')2 fragment specific, AP conjugate. 1:5000 diluted). A fluorescent signal was created by adding the AP substrate Attophos. The signal on the antigen is at least 5-fold above background, whereas the signal on the control antigens is less than 1.5-fold above background.
Application
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
ELISA
2ug/ml
Immunohistology - Frozen
2ug/ml
Western Blotting
2ug/ml
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Western Blotting
HCA008 detects a band of approximately 191 kDa in mouse lung cell lysates.
Technical Advice
Protocols and further information may be found in the HuCALTechnical Manual.
Storage
Prior to reconstitution store at +4oC.
After reconstitution store at -20oC.
Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
6 months from date of reconstitution.
Licensed Use
For in vitro research purposes only, unless otherwise specified in writing by AbD Serotec.
Sold under license of U.S. Patents 6,300,064, 6,696,248, 6,708,484, 6,753,136, European Patent 0,859,841 and corresponding patents.
This antibody was developed byAbD Serotec, a wholly owned subsidiary of Bio-Rad Laboratories Inc., Zeppelinstr. 4, 82178 Puchheim, Germany, in collaboration with theKazusa DNA Research Institute, 2-6-7 Kazusa-kamatari, Kisarazu, Chiba 292-0818 JAPAN.
WB data kindly provided by the Kazusa DNA Research Institute, Japan.
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