CH25H ELISA Kit (Human) (OKCD01912)

Catalog No: OKCD01912 (Formerly GWB-KBBCI3)

Size:96WELLS

Human CH25H ELISA Kit Standard Curve

Product Info

• Predicted Species Reactivity: Homo sapiens|Human
• Application: Enzyme-linked Immunosorbent assay-Sandwich
• ELISA Kit Detection Method: Colorimetric
• ELISA Kit Duration: 3h
• ELISA Kit Linearity:

  Sample	1:2 Dilution	1:4 Dilution	1:8 Dilution	1:16 Dilution
  Serum (n=5)	99-105%	94-102%	97-104%	78-97%
  EDTA Plasma (n=5)	80-96%	87-101%	88-95%	78-104%
  Heparin Plasma (n=5)	82-101%	80-98%	90-97%	89-103%

• ELISA Kit 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.
• ELISA Kit Range: 125-8000pg/mL
• ELISA Kit Recovery:

  Matrix	Recovery Range (%)	Average (%)
  Serum (n=5)	99-105	102
  EDTA Plasma (n=5)	92-103	99
  Heparin Plasma (n=5)	83-95	86

• ELISA Kit 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%
• ELISA Kit Component:
  • Anti-CH25H Microplate: 96 Wells (12 x 8 Well Strips)
  • CH25H Lyophilized Standard: 2 x 8 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
• Reconstitution and Storage: 2°C to 8°C|-20°C
• Sample Type: Serum, plasma, tissue homogenates and other biological fluids
• Sensitivity: < 49 pg/mL
• 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.
• Assay Info: Assay Methodology: Quantitative Sandwich Immunoassay

Target Info

• Gene Symbol: CH25H
• Gene Full Name: cholesterol 25-hydroxylase
• Alias Symbols: C25H;cholesterol 25-hydroxylase;cholesterol 25-monooxygenase;h25OH.
• NCBI Gene Id: 9023
• Protein Name: Cholesterol 25-hydroxylase
• Description of 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. May play an important role in regulating lipid metabolism by synthesizing a corepressor that blocks sterol regulatory element binding protein (SREBP) processing. In testis, production of 25-hydroxycholesterol by macrophages may play a role in Leydig cell differentiation.
• Uniprot ID: O95992
• Protein Accession #: NP_003947.1
• Nucleotide Accession #: NM_003956.3
• Protein Size (# AA): 272