TSG101 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Tumor Susceptibility Gene 101 (TSG101) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Tumor Susceptibility Gene 101 (TSG101) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

ESCRT-I complex subunit TSG101; TSG10; tumor susceptibility gene 10; tumor susceptibility gene 101; tumor susceptibility gene 101 protein; tumor susceptibility protein; VPS23.

Component of the ESCRT-I complex, a regulator of vesicular trafficking process. Binds to ubiquitinated cargo proteins and is required for the sorting of endocytic ubiquitinated cargos into multivesicular bodies (MVBs) . Mediates the association between the ESCRT-0 and ESCRT-I complex. Required for completion of cytokinesis; the function requires CEP55. May be involved in cell growth and differentiation. Acts as a negative growth regulator. Involved in the budding of many viruses through an interaction with viral proteins that contain a late-budding motif P-[ST]-A-P. This interaction is essential for viral particle budding of numerous retroviruses. Required for the exosomal release of SDCBP, CD63 and syndecan (PubMed:22660413) . It may also play a role in the extracellular release of microvesicles that differ from the exosomes (PubMed:22315426) .

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 Tumor Susceptibility Gene 101 (TSG101) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Tumor Susceptibility Gene 101 (TSG101) . 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 Tumor Susceptibility Gene 101 (TSG101), 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 Tumor Susceptibility Gene 101 (TSG101) 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) 78-98 91 EDTA Plasma (n=5) 86-101 93 Heparin Plasma (n=5) 78-88 83 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 78-98 91 Serum (n=5) 78-98 91 EDTA Plasma (n=5) 86-101 93 EDTA Plasma (n=5) 86-101 93 Heparin Plasma (n=5) 78-88 83 Heparin Plasma (n=5) 78-88 83

Component Amount Anti-TSG101 Microplate 96 Wells (12 x 8 Well Strips) TSG101 Lyophilized Standard 2 x 20 ng 100X Biotinylated TSG101 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-TSG101 Microplate 96 Wells (12 x 8 Well Strips) Anti-TSG101 Microplate 96 Wells (12 x 8 Well Strips) TSG101 Lyophilized Standard 2 x 20 ng TSG101 Lyophilized Standard 2 x 20 ng 100X Biotinylated TSG101 Detector Antibody 120 uL 100X Biotinylated TSG101 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 Tumor Susceptibility Gene 101 (TSG101) . No significant cross-reactivity or interference between Tumor Susceptibility Gene 101 (TSG101) and analogues was observed.