Description

Nori Feline TNF RII ELISA Kit Summary

Alternative names for TNF RII: tumor necrosis factor receptor II, CD120b, TNFRII, TNFRSF1B, death receptor, TNFR2

Alternative names for feline: cat

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background:

A tumor necrosis factor receptor (TNFR), or death receptor, is a trimeric^[2] cytokine receptor that binds tumor necrosis factors (TNF).^[1] The receptor cooperates with an adaptor protein (such as TRADD, TRAF, RIP), which is important in determining the outcome of the response (e.g. apoptosis, inflammation). Because “TNF” is often used to describe TNF alpha, “TNFR” is often used to describe the receptors that bind to TNF alpha – namely, CD120. However, there are several other members of this family that bind to the other TNFs.^[3][4] While all cell types studied express TNF RI, TNF RII expression is limited primarily to hematopoietic cells and cells of the immune system. Most of the biological function of TNF are mediated via TNF RI. Soluble forms of both TNF RI and TNF RII, generated as a result of proteolytic cleavage of the extracellular domain, have been detected in vivo in various fluids. The soluble TNF receptors can bind TNF with high affinity and function as TNF-antagonists. ^{[3, 5-7]}

Reference

1. Banner DW, D’Arcy A, Janes W et al. (May 1993). Cell 73 (3): 431–45.
2. Ashkenazi, A.; Dixit, VM (1998). Science 281 (5381): 1305–8.
3. Locksley RM, Killeen N, Lenardo MJ (2001). Cell 104 (4): 487–501.
4. Hehlgans T, Pfeffer K (2005). Immunology 115 (1): 1–20.
5. Chen G & DV Goeddel (2002) Science 296: 1634.
6. MacEwan DJ (2002) Cell Signal 14: 477.
7. Feldmann M & RN Maini (2001) Annu. Rev. Immunol. 19:163.

Product Citations