TRAF3 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level TNF Receptor Associated Factor 3 (TRAF3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level TNF Receptor Associated Factor 3 (TRAF3) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

CAP1; CAP-1; CD40 associated protein 1; CD40 binding protein; CD40 receptor associated factor 1; CD40 receptor-associated factor 1; CD40-binding protein; CD40bp; CRAF1; IIAE5; LAP1; LMP1-associated protein 1; RING-type E3 ubiquitin transferase TRAF3; RNF118; TNF receptor-associated factor 3.

Regulates pathways leading to the activation of NF-kappa-B and MAP kinases, and plays a central role in the regulation of B-cell survival. Part of signaling pathways leading to the production of cytokines and interferon. Required for normal antibody isotype switching from IgM to IgG. Plays a role T-cell dependent immune responses. Plays a role in the regulation of antiviral responses. Is an essential constituent of several E3 ubiquitin-protein ligase complexes. May have E3 ubiquitin-protein ligase activity and promote 'Lys-63'-linked ubiquitination of target proteins. Inhibits activation of NF-kappa-B in response to LTBR stimulation. Inhibits TRAF2-mediated activation of NF-kappa-B. Down-regulates proteolytic processing of NFKB2, and thereby inhibits non-canonical activation of NF-kappa-B. Promotes ubiquitination and proteasomal degradation of MAP3K14.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to TNF Receptor Associated Factor 3 (TRAF3) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to TNF Receptor Associated Factor 3 (TRAF3) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain TNF Receptor Associated Factor 3 (TRAF3), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of TNF Receptor Associated Factor 3 (TRAF3) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) Serum (n=5) 79-103 89 EDTA Plasma (n=5) 85-92 88 Heparin Plasma (n=5) 86-95 90 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 79-103 89 Serum (n=5) 79-103 89 EDTA Plasma (n=5) 85-92 88 EDTA Plasma (n=5) 85-92 88 Heparin Plasma (n=5) 86-95 90 Heparin Plasma (n=5) 86-95 90

Component Amount Anti-TRAF3 Microplate 96 Wells (12 x 8 Well Strips) TRAF3 Lyophilized Standard 2 x 1 ng 100X Biotinylated TRAF3 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-TRAF3 Microplate 96 Wells (12 x 8 Well Strips) Anti-TRAF3 Microplate 96 Wells (12 x 8 Well Strips) TRAF3 Lyophilized Standard 2 x 1 ng TRAF3 Lyophilized Standard 2 x 1 ng 100X Biotinylated TRAF3 Detector Antibody 120 uL 100X Biotinylated TRAF3 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of TNF Receptor Associated Factor 3 (TRAF3) . No significant cross-reactivity or interference between TNF Receptor Associated Factor 3 (TRAF3) and analogues was observed.