SKP1 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level S-Phase Kinase Associated Protein 1 (SKP1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level S-Phase Kinase Associated Protein 1 (SKP1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Cyclin A/CDK2-associated p19; cyclin-A/CDK2-associated protein p19; EMC19; OCP2; OCP-2; OCP-II; organ of Corti protein 2; organ of Corti protein II; p19A; p19skp1; RNA polymerase II elongation factor-like protein; RNA polymerase II elongation factor-like protein OCP2; SIII; SKP1A; S-phase kinase-associated protein 1; TCEB1L; transcription elongation factor B polypeptide 1-like.

Essential component of the SCF (SKP1-CUL1-F-box protein) ubiquitin ligase complex, which mediates the ubiquitination of proteins involved in cell cycle progression, signal transduction and transcription. In the SCF complex, serves as an adapter that links the F-box protein to CUL1. The functional specificity of the SCF complex depends on the F-box protein as substrate recognition component. SCF (BTRC) and SCF (FBXW11) direct ubiquitination of CTNNB1 and participate in Wnt signaling. SCF (FBXW11) directs ubiquitination of phosphorylated NFKBIA. SCF (BTRC) directs ubiquitination of NFKBIB, NFKBIE, ATF4, SMAD3, SMAD4, CDC25A, FBXO5, CEP68 and probably NFKB2 (PubMed:25704143) . SCF (SKP2) directs ubiquitination of phosphorylated CDKN1B/p27kip and is involved in regulation of G1/S transition. SCF (SKP2) directs ubiquitination of ORC1, CDT1, RBL2, ELF4, CDKN1A, RAG2, FOXO1A, and probably MYC and TAL1. SCF (FBXW7) directs ubiquitination of cyclin E, NOTCH1 released notch intracellular domain (NICD), and probably PSEN1. SCF (FBXW2) directs ubiquitination of GCM1. SCF (FBXO32) directs ubiquitination of MYOD1. SCF (FBXO7) directs ubiquitination of BIRC2 and DLGAP5. SCF (FBXO33) directs ubiquitination of YBX1. SCF (FBXO11) directs ubiquitination of BCL6 and DTL but does not seem to direct ubiquitination of TP53. SCF (BTRC) mediates the ubiquitination of NFKBIA at 'Lys-21' and 'Lys-22'; the degradation frees the associated NFKB1-RELA dimer to translocate into the nucleus and to activate transcription. SCF (CCNF) directs ubiquitination of CCP110. SCF (FBXL3) and SCF (FBXL21) direct ubiquitination of CRY1 and CRY2. SCF (FBXO9) directs ubiquitination of TTI1 and TELO2. SCF (FBXO10) directs ubiquitination of BCL2.

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 S-Phase Kinase Associated Protein 1 (SKP1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to S-Phase Kinase Associated Protein 1 (SKP1) . 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 S-Phase Kinase Associated Protein 1 (SKP1), 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 S-Phase Kinase Associated Protein 1 (SKP1) 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 Anti-SKP1 Microplate 96 Wells (12 x 8 Well Strips) SKP1 Lyophilized Standard 2 x 10 ng 100X Biotinylated SKP1 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-SKP1 Microplate 96 Wells (12 x 8 Well Strips) Anti-SKP1 Microplate 96 Wells (12 x 8 Well Strips) SKP1 Lyophilized Standard 2 x 10 ng SKP1 Lyophilized Standard 2 x 10 ng 100X Biotinylated SKP1 Detector Antibody 120 uL 100X Biotinylated SKP1 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 S-Phase Kinase Associated Protein 1 (SKP1) . No significant cross-reactivity or interference between S-Phase Kinase Associated Protein 1 (SKP1) and analogues was observed.