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Recombinant Human Cochlin Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Bioactivity
Format
Carrier-Free

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Recombinant Human Cochlin Protein, CF Summary

Details of Functionality
Measured by the ability of the immobilized protein to support the adhesion of SW1353 human chondrosarcoma cells. When 5 x 104 cells per well are added to rhCochlin coated plates (10 μg/mL, 100 μL/well) approximately 70-90% will adhere after 60 minutes at 37 °C.
Source
Mouse myeloma cell line, NS0-derived human Cochlin protein
Glu25-Gln550, with an N-terminal 6-His tag
Accession #
N-terminal Sequence
His
Protein/Peptide Type
Recombinant Proteins
Gene
COCH
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Note
<0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Bioactivity
Theoretical MW
58 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
64-66 kDa, reducing conditions

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Buffer
Lyophilized from a 0.2 μm filtered solution in PBS.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Reconstitution Instructions
Reconstitute at 100 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Cochlin Protein, CF

  • coagulation factor C (Limulus polyphemus homolog); cochlin
  • coagulation factor C homolog, cochlin (Limulus polyphemus)
  • COCH
  • COCH5B2
  • COCH-5B2COCH5B2
  • Cochlin
  • DFNA31
  • DFNA9

Background

Cochlin, also known as Coch-5B2, is an N-glycosylated protein that constitutes the major protein component of inner ear extracellular matrix. Human Cochlin contains one LCCL/FCH and two VWF-A domains, all three of which may be deleted by alternate splicing (1 - 3). The predominant species in human, mouse, rat, and cow are the 63 kDa full length protein, two 40 and 46 kDa isoforms that lack the LCCL domain, and a 16 kDa isoform that lacks both VWF-A domains (3, 4). Cochlin is also susceptible to proteolysis between the LCCL and VWF-A1 domains (5). Mature full length human Cochlin shares 96% amino acid sequence identity with bovine, mouse, and rat Cochlin. Cochlin is secreted by inner ear fibrocytes and accumulates in acidophilic deposits of the cochlea and vestibule (6 - 10). Several point mutations in the LCCL domain are associated with the autosomal hearing loss disorder DFNA9, and the resulting mutant proteins exhibit altered aggregation and matrix association properties (2, 7, 11). The 63 and 16 kDa isoforms are also present in perilymph fluid (4). Cochlin is a target of IFN-gamma producing T cells in autoimmune sensorineural hearing loss (ASNHL) (12). Cochlin associates with the choline transporter CTL2 which itself is a target of autoimmune-mediated hearing loss (3, 13). Cochlin, absent in normal trabecular meshwork (TM) of the eye, is upregulated in glaucoma and is deposited around TM cells. It promotes the in vitro aggregation of TM cells and TM cell adhesion to collagen (14, 15). Polymorphisms within the second VFW-A domain are associated with glaucoma (15).

  1. Aggarwal, S. and A.L. Gurney (2002) J. Leukoc. Biol. 71:1.
  2. Moseley, T.A. et al. (2003) Cytokine & Growth Factor Rev. 14:155.
  3. Hymowitz, S.G. et al. (2001) EMBO J. 20:5332.
  4. Haudenschild, D. et al. (2002) J. Biol. Chem. 277:4309.
  5. Starnes, T. et al. (2002) J. Immunol. 169:642.
  6. Li, H. et al. (2000) Proc. Natl. Acad. Sci. USA 97:773.
  7. Robertson, N.G. et al. (2006) Hum. Mol. Genet. 15:1071.
  8. Mizuta, K. et al. (2008) Neurosci. Lett. 434:104.
  9. Li, L. et al. (2005) Auris Nasus Larynx 32:219.
  10. Shindo, S. et al. (2008) Neurosci. Lett. 444:148.
  11. Grabski, R. et al. (2003) Hum. Genet. 113:406.
  12. Baek, M.-J. et al. (2006) J. Immunol. 177:4203.
  13. Nair, T.S. et al. (2004) J. Neurosci. 24:1772.
  14. Bhattacharya, S.K. et al. (2005) J. Biol. Chem. 280:6080.
  15. Picciani, R. et al. (2007) Prog. Retin. Eye Res. 26:453.
 

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Bioinformatics

Gene Symbol COCH
Uniprot