OPG/OCIF

Specification sheet	Human OPG/OCIF ELISA Kit
Catalog No: OCK002-S1	One plate; Stripe; 96well determinations
Catalog No: OCK002-C3	Three plate; Complete; 288well determinations
Application	Human OPG/OCIF is a capture ELISA for the quantitative measurement of human OPG/OCIF in culture media and serum
Sensitivity	30pg/ml	Range of Detection	30-200pg/ml
Specificity	Native and recombinant human OPG/OCIF
Assay time	Total assay time is 5 - 6hrs at room temperature
Storage	Store at 2-8^oC	Stability	5 months when stored at 4^oC

Kit Reagents Supplied

1.anti-OPG/OCIF precoated microplate	2.Lyophilized rhOPG/OCIF standard
3.Streptavidin-HRP concentrate	4.Biotinylated antibody reagent
5.Streptavidin-HRP dilution buffer	6.Stop solution, 0.18M phosphoric acid
7.Premixed TMB substrate solution A	8.Premixed TMB substrate solution B
9.Wash buffer 30-fold concentrated	10.Adhesive plate covers	11.Standard diluent

Protocol ( For Research Use Only )
1. Dispense 100µl of the standards, controls, and samples into their designated wells.
2. Seal the plate and incubate for 2hours at the room temperature.
3. Wash the plate 3 times with Wash buffer.
4. Add 100µl of the diluted Biotinylated- antibody to each well.
5. Seal the plate and incubate it for 2hours at the room temperature.
6. Wash the plate 3 times with wash buffer.
7. Add 100µl of the diluted Streptavidin-HRP conjugate to each well.
8. Seal the plate and incubate it for 30min at the room temperature.
9. Wash the plate more than 4 times with wash buffer.
10. Immediately dispense 100µl TMB substrate solution (TMB A + TMB B) into each well and incubate at room temperature for 15-30mins.
11. Add 50µl of Stop Solution to each well.
12. Read absorbance with a microplate reader at 450nm.
Reference
Simonet W.S. et al.: Osteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell 89, 309-319 (1997)
Yasuda H., Shima N., Nakagawa N. et al.: Identity of osteoclastogenesis inhibitory factor (OCIF) and osteoprotegerin (OPG) : a mechanism by which OPG/OCIF inhibits osteogenesis in vitroEndocrinology 139, 1329-1337 (1998)