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Ch25h ELISA Kit (Mouse)

CAT: 0247-OKCD01913-96WSize: 96 WellDry Ice: NoHazardous: No
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CAT#:0247-OKCD01913-96WSize:96 Well
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24/48H Stock Items & 2 to 6 Weeks non Stock Items.
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Reproducibility
Intra-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%
CAS Number
7732-18-5
Gene Name
Cholesterol 25-hydroxylase
Gene Aliases
AI462618; cholesterol 25-hydroxylase; cholesterol 25-monooxygenase; m25O; m25OH.
Gene ID
12642
Accession Number
NP_034020.1
Reactivity
Mouse|Mus musculus
Target
Catalyzes 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) .
Type
ELISA Kit
Applications
Enzyme-linked Immunosorbent assay-Sandwich
Detection Method
Colorimetric
Assay Principle
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 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.
Assay Protocol
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
Sample Type
Tissue homogenates, cell lysates and other biological fluids
Detection Range
0.78-50ng/mL
Sensitivity
< 0.27 ng/mL
Reconstitution
2°C to 8°C|-20°C
Components
Component Amount Anti-Ch25h Microplate 96 Wells (12 x 8 Well Strips) Ch25h Lyophilized Standard 2 x 50 ng 100X Biotinylated Ch25h 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-Ch25h Microplate 96 Wells (12 x 8 Well Strips) Anti-Ch25h Microplate 96 Wells (12 x 8 Well Strips) Ch25h Lyophilized Standard 2 x 50 ng Ch25h Lyophilized Standard 2 x 50 ng 100X Biotinylated Ch25h Detector Antibody 120 uL 100X Biotinylated Ch25h 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
Notes
Formerly GWB-KBBJA5
Specificity
This 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.
Protein Length
298
NCBI Gene Symbol
Ch25h
Protein Name
Cholesterol 25-hydroxylase
Gene Name URL
Ch25h
Nucleotide Accession Number
NM_009890.1

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