Specification sheet

Human TNF- α ELISA Kit
Catalog No: OCK001-S1 One plate; Stripe; 96well determinations
Catalog No: OCK001-C3 Three plate;  Complete; 288well determinations
Application

Human TNF-α is a capture ELISA for the quantitative measurement of human TNF-α in culture media and serum
Sensitivity 2pg/ml Range of Detection 15-1500pg/ml
Specificity Native and recombinant human TNF-α Storage Store at 2-8oC
Assay time Total assay time is 5 - 6hrs at room temperature Stability 5 months when stored at 4oC

Kit Reagents Supplied
1.anti-hTNF-α precoated microplate 2.Lyophilized rhTNF-a 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. click to see Contents...
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
Avirutnan, P., P. Malasit, B. Seliger, S. Bhakdi, and M. Husmann (1998) Dengue virus infection of human endothelial cells leads to chemokine production, complement activation, and apoptosis. J.Immunol 161(11):6338-46.
Aziz, N., et al.  (1999)  Stability of Plasma Levels of Cytokines and Soluble Activation Markers in Patients with Human Immunodeficiency Virus Infection. Journal of Infectious Disease. 179:843-8.
Bienvenu, J., et al.  (1993)  Analytical performances of commercial ELISA kits for IL-2, IL-6 and TNF-alpha.  A WHO study.  Eur. Cytokine Netw.  4(6);447-451.
Oliver, S.J., A. Moreira, and G. Kaplan (2000) Immune stimulation in scleroderma patients treated with thalidomide. Clinical Immunology 97(2):1980120.