Insulin-like 3, Recombinant, Human (INSL3, Leydig Cell Insulin-like, Ley I-L)

Human INSL3 (insulin-like 3), formerly called Ley I-L (Leydig cell insulin-like), is a member of the insulin/relaxin superfamily.1-3 INSL3 is a secreted protein produced in high amounts by testicular Leydig cells, and in lower amounts by ovarian follicular theca interna cells.3, 4 As with other insulin/relaxin superfamily members, human INSL3 is synthesized as a preprohormone.1 The PreproINSL3 precursor is 131 amino acids (aa) in length. It is processed by a prohormone convertase to remove a 20 aa signal sequence and a 47 aa internal pro-, or C-peptide that separates a 35 aa N-terminal B chain from a 26 aa C-terminal A chain. The resulting mature protein is an unglycosylated 6kD complex of disulfide-linked A and B chains.1-3 The proteolytic processing site(s) for some species is uncertain. Based on human, mature human INSL3 shares 70%, 62%, 67% and 62% aa identity with canine, rat, porcine and mouse INSL3, respectively. The receptor for INSL3 is the leucine-rich G-protein coupled receptor LGR8, also called RXFP2 or GREAT.5 LRG8 is mainly expressed near the sites of INSL3 secretion, but also in brain, embryonic kidney and other tissues.1 Deletion of either INSL3 or LGR8 in the mouse causes failure of the testes to descend (cryptorchidism), due to an altered development of the gubernaculum, which is a cord that connects the fetal testes to the bottom of the scrotum.5-8 Mutation of INSL3 is also an occasional cause of cryptorchidism in humans.9 In the male, INSL3 increases in the circulation during puberty, reaching a maximum in the adult.10, 11 In the female, circulating INSL3 correlates with follicle development.3, 4, 12 For both, luteinizing hormone (LH) is the major stimulus of INSL3 expression.10, 11\n\nSource: A DNA sequence encoding human INSL3 (Leu 21-Try 131; Accession # P51460) was expressed in E. coli.\n\nDefinition: Measured by its ability to induce cAMP accumulation in human RXFP2 transfected HEK293 cells.\n\nApplications: \nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: The A chain has a calculated molecular mass of ~ 2.8kD; the B chain has a calculated molecular mass of ~ 3.8kD.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nLyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile PBS and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

I7660-85

Size

50ug

Applications

WB

Reactivities

Hum

Form

Supplied as a lyophilized powder in PBS. Reconstitute with sterile PBS to (same/more than) 0.1mg/ml.

Purity

(same/more than) 95%, as determined by SDS-PAGE and visualized by silver stain. Endotoxin (same/less than) 1EU.

References

1. Bathgate, R.A. et al. (2006) Pharmacol. Rev. 58:7. 7. Nef, S. L.F. Parada (1999) Nat. Genet. 22:295.\n2. Burkhardt, E. et al. (1994) Hum. Genet. 94:91. 8. Gorlov, I.P. et al. (2002) Hum. Mol. Genet. 11:2309.\n3. Zimmerman, S. et al. (1997) Mol. Reprod. Dev. 47:30. 9. Tomboc, M. et al. (2000) J. Clin. Endocrinol. Metab. 85:4013.\n4. Tashima, L.S. et al. (1995) J. Clin. Endocrinol. Metab. 80:707. 10. Foresta, C. et al. (2004) J. Clin. Endocrinol. Metab. 89:5952.\n5. Kumagai, J. et al. (2002) J. Biol. Chem. 277:31283. 11. Ferlin, A. et al. (2006) J. Clin. Endocrinol. Metab. 91:3426.\n6. Zimmerman, S. et al. (1999) Mol. Endocrinol. 13:681.