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Catalog No: OKCD01913 (Formerly GWB-KBBJA5)
Size:96WELLS
Price: $700.00
SKU
OKCD01913
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Datasheets/ManualsPrintable datasheet for Ch25h ELISA Kit (Mouse) (OKCD01913)
Product Info
Predicted Species ReactivityMouse|Mus musculus
ApplicationEnzyme-linked Immunosorbent assay-Sandwich
ELISA Kit Detection MethodColorimetric
ELISA Kit Duration3h
ELISA Kit PrincipleThe 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 Cholesterol-25-Hydroxylase (CH25H). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Cholesterol-25-Hydroxylase (CH25H). 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 Cholesterol-25-Hydroxylase (CH25H), 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 Cholesterol-25-Hydroxylase (CH25H) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
ELISA Kit Range0.78-50ng/mL
ELISA Kit ReproducibilityIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cholesterol-25-Hydroxylase (CH25H) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cholesterol-25-Hydroxylase (CH25H) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
ELISA Kit Component
ComponentAmount
Anti-Ch25h Microplate96 Wells (12 x 8 Well Strips)
Ch25h Lyophilized Standard2 x 50 ng
100X Biotinylated Ch25h Detector Antibody120 uL
Avidin/HRP Conjugate120 uL
Standard Diluent1 x 20 mL
Detector Antibody Diluent1 x 12 mL
Conjugate Diluent1 x 12 mL
30X Wash Buffer1 x 20 mL
TMB Substrate1 x 9 mL
Stop Solution1 x 6 mL
Reconstitution and Storage2°C to 8°C|-20°C
Sample TypeTissue homogenates, cell lysates and other biological fluids
Sensitivity< 0.27 ng/mL
SpecificityThis assay has high sensitivity and excellent specificity for detection of Cholesterol-25-Hydroxylase (CH25H). No significant cross-reactivity or interference between Cholesterol-25-Hydroxylase (CH25H) and analogues was observed.
Assay InfoAssay Methodology: Quantitative Sandwich Immunoassay
Gene SymbolCh25h
Gene Full Namecholesterol 25-hydroxylase
Alias SymbolsAI462618;cholesterol 25-hydroxylase;cholesterol 25-monooxygenase;m25O;m25OH.
NCBI Gene Id12642
Protein NameCholesterol 25-hydroxylase
Description of TargetCatalyzes the formation of 25-hydroxycholesterol from cholesterol, leading to repress cholesterol biosynthetic enzymes (PubMed:9852097). Plays a key role in cell positioning and movement in lymphoid tissues: 25-hydroxycholesterol is an intermediate in biosynthesis of 7-alpha,25-dihydroxycholesterol (7-alpha,25-OHC), an oxysterol that acts as a ligand for the G protein-coupled receptor GPR183/EBI2, a chemotactic receptor for a number of lymphoid cells (PubMed:22999953). May play an important role in regulating lipid metabolism by synthesizing a corepressor that blocks sterol regulatory element binding protein (SREBP) processing (PubMed:9852097). In testis, production of 25-hydroxycholesterol by macrophages may play a role in Leydig cell differentiation (PubMed:9852097).
Uniprot IDQ9Z0F5
Protein Accession #NP_034020.1
Nucleotide Accession #NM_009890.1
Protein Size (# AA)298
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