RIPK1 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Receptor Interacting Serine Threonine Kinase 1 (RIPK1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Receptor Interacting Serine Threonine Kinase 1 (RIPK1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

AIEFL; cell death protein RIP; IMD57; receptor (TNFRSF) -interacting serine-threonine kinase 1; receptor-interacting protein 1; receptor-interacting protein kinase 1; receptor-interacting serine/threonine-protein kinase 1; RIP; RIP1; RIP-1; serine/threonine-protein kinase RIP.

Serine-threonine kinase which transduces inflammatory and cell-death signals (programmed necrosis) following death receptors ligation, activation of pathogen recognition receptors (PRRs), and DNA damage. Upon activation of TNFR1 by the TNF-alpha family cytokines, TRADD and TRAF2 are recruited to the receptor. Phosphorylates DAB2IP at 'Ser-728' in a TNF-alpha-dependent manner, and thereby activates the MAP3K5-JNK apoptotic cascade. Ubiquitination by TRAF2 via 'Lys-63'-link chains acts as a critical enhancer of communication with downstream signal transducers in the mitogen-activated protein kinase pathway and the NF-kappa-B pathway, which in turn mediate downstream events including the activation of genes encoding inflammatory molecules. Polyubiquitinated protein binds to IKBKG/NEMO, the regulatory subunit of the IKK complex, a critical event for NF-kappa-B activation. Interaction with other cellular RHIM-containing adapters initiates gene activation and cell death. RIPK1 and RIPK3 association, in particular, forms a necrosis-inducing complex.

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 Receptor Interacting Serine Threonine Kinase 1 (RIPK1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Receptor Interacting Serine Threonine Kinase 1 (RIPK1) . 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 Receptor Interacting Serine Threonine Kinase 1 (RIPK1), 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 Receptor Interacting Serine Threonine Kinase 1 (RIPK1) 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

Component Amount _x005F Anti-RIPK1 Microplate 96 Wells (12 x 8 Well Strips) _x005F RIPK1 Lyophilized Standard 2 x 10 ng _x005F 100X Biotinylated RIPK1 Detector Antibody 120 uL _x005F Avidin/HRP Conjugate 120 uL _x005F Standard Diluent 1 x 20 mL _x005F Detector Antibody Diluent 1 x 12 mL _x005F Conjugate Diluent 1 x 12 mL _x005F 30X Wash Buffer 1 x 20 mL _x005F TMB Substrate 1 x 9 mL _x005F Stop Solution 1 x 6 mL _x005F Component Amount Component Amount Anti-RIPK1 Microplate 96 Wells (12 x 8 Well Strips) Anti-RIPK1 Microplate 96 Wells (12 x 8 Well Strips) RIPK1 Lyophilized Standard 2 x 10 ng RIPK1 Lyophilized Standard 2 x 10 ng 100X Biotinylated RIPK1 Detector Antibody 120 uL 100X Biotinylated RIPK1 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 Receptor Interacting Serine Threonine Kinase 1 (RIPK1) ._x005F No significant cross-reactivity or interference between Receptor Interacting Serine Threonine Kinase 1 (RIPK1) and analogues was observed.