Sumo1 ELISA Kit (Mouse)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Small Ubiquitin Related Modifier Protein 1 (SUMO1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Small Ubiquitin Related Modifier Protein 1 (SUMO1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

GMP1; PI; PIC1; SE; SENTRIN; small ubiquitin-related modifier 1; SMT3; SMT3 homolog 3; SMT3 suppressor of mif two 3 homolog 1; Smt3C; SMT3H3; SMTP3; SUMO-1; SUMO-1 related peptidase; ubiquitin-homology domain protein PIC1; ubiquitin-like 1; ubiquitin-like protein SMT3C; Ubl; Ubl1.

Ubiquitin-like protein that can be covalently attached to proteins as a monomer or a lysine-linked polymer. Covalent attachment via an isopeptide bond to its substrates requires prior activation by the E1 complex SAE1-SAE2 and linkage to the E2 enzyme UBE2I, and can be promoted by E3 ligases such as PIAS1-4, RANBP2 or CBX4. This post-translational modification on lysine residues of proteins plays a crucial role in a number of cellular processes such as nuclear transport, DNA replication and repair, mitosis and signal transduction. Involved for instance in targeting RANGAP1 to the nuclear pore complex protein RANBP2. Covalently attached to the voltage-gated potassium channel KCNB1; this modulates the gating characteristics of KCNB1. Polymeric SUMO1 chains are also susceptible to polyubiquitination which functions as a signal for proteasomal degradation of modified proteins. May also regulate a network of genes involved in palate development. Covalently attached to ZFHX3 (By similarity) .

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 Small Ubiquitin Related Modifier Protein 1 (SUMO1) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Small Ubiquitin Related Modifier Protein 1 (SUMO1) . 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 Small Ubiquitin Related Modifier Protein 1 (SUMO1), 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 Small Ubiquitin Related Modifier Protein 1 (SUMO1) 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) 90-97 93 EDTA Plasma (n=5) 78-96 86 Heparin Plasma (n=5) 80-97 94 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 90-97 93 Serum (n=5) 90-97 93 EDTA Plasma (n=5) 78-96 86 EDTA Plasma (n=5) 78-96 86 Heparin Plasma (n=5) 80-97 94 Heparin Plasma (n=5) 80-97 94

Component Amount Anti-Sumo1 Microplate 96 Wells (12 x 8 Well Strips) Sumo1 Lyophilized Standard 2 x 10 ng 100X Biotinylated Sumo1 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-Sumo1 Microplate 96 Wells (12 x 8 Well Strips) Anti-Sumo1 Microplate 96 Wells (12 x 8 Well Strips) Sumo1 Lyophilized Standard 2 x 10 ng Sumo1 Lyophilized Standard 2 x 10 ng 100X Biotinylated Sumo1 Detector Antibody 120 uL 100X Biotinylated Sumo1 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 Small Ubiquitin Related Modifier Protein 1 (SUMO1) . No significant cross-reactivity or interference between Small Ubiquitin Related Modifier Protein 1 (SUMO1) and analogues was observed.