Recombinant Human Cochlin Protein, CF

Catalog # Availability Size / Price Qty
5249-CH-050
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Recombinant Human Cochlin Protein, CF Summary

Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
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
Analysis
His
Predicted Molecular Mass
58 kDa
SDS-PAGE
64-66 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

5249-CH

Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.
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Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Cochlin

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).

References
  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.
 
Entrez Gene IDs
1690 (Human); 12810 (Mouse); 362735 (Rat)
Alternate Names
coagulation factor C (Limulus polyphemus homolog); cochlin; coagulation factor C homolog, cochlin (Limulus polyphemus); COCH; COCH5B2; COCH-5B2COCH5B2; Cochlin; DFNA31; DFNA9

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