HCA010 binds to the C-terminal 82 amino acids of the mKIAA0531 protein, also known as kinesin family member 5C, part of neuron-specific molecular motors acting in the microtubule-based motilities of organelles. Kinesins are force-producing proteins that may play a role in organelle transport.
It is composed of three structural domains: a large globular N-terminal domain which is responsible for the motor activity of kinesin (it hydrolyzes ATP and binds microtubule), a central alpha-helical coiled coil domain that mediates the heavy chain dimerization; and a small globular c-terminal domain which interacts with other proteins (such as the kinesin light chains), vesicles and membranous organelles.
Due to high homology, HCA010 may cross-react with the other members of the kinesin family 5 (KIF5A, KIF5B) as well as with the kinesin heavy chains.
HCA010 may also cross-react with human kinesin family member 5C (KIAA0531) as there is 97% identity with the antigen.
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 mKIAA0531 (5 µg/ml) plus unrelated control proteins were immobilized on a microtiter plate. Specific binding was monitored by first adding HCA010 (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
Western Blotting
0.5ug/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
HCA010 detects a band of approximately 113 kDa in mouse lung cell lysates.
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.
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