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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 2-8oC Stability 5 months when stored at 4oC

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.

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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)
 

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