CH25H ELISA Kit
Quick Overview for CH25H ELISA Kit (ABIN5652905)
Target
See all CH25H ELISA KitsReactivity
Detection Method
Method Type
Detection Range
Application
Sample Type
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Minimum Detection Limit
- 0.78 ng/mL
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Purpose
- Mouse Cholesterol-25-Hydroxylase (CH25H) ELISA Kit for detecting CH25H in tissue homogenates, cell lysates
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Analytical Method
- Quantitative
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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.
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Sensitivity
- 0.29 ng/mL
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Components
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- Pre-coated, ready to use 96-well strip plate
- Standar
- Detection Reagent A
- Detection Reagent B
- TMB Substrate
- Stop Solution
- Assay Diluent A
- Assay Diluent B
- Wash Buffer (30x concentrate)
- Standard Diluent
- Plate Sealer
- Instruction Manual
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Material not included
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- Microplate Reader with 450 nm filter
- Single or multi-channel pipettes with high precision and disposable tips
- Microcentrifuge tubes
- Deionized or distilled watter
- Absorbent paper for blotting the microplate
- Container for wash solution
- 0.01 M (or 1x) Phosphate Buffered Saline (PBS), ph 7.0-7.2
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Application Notes
- Optimal working dilution should be determined by the investigator.
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Comment
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This product is tested and assembled before shipping to ensure that it meets its quality specifications. All kits are tested to confirm that they fall within their defined Inter- and Intra- assay coefficient of variation.
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Sample Volume
- 100 μL
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Assay Time
- 3 h
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Plate
- Pre-coated
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Protocol
- 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.
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Assay Precision
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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% -
Restrictions
- For Research Use only
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Handling Advice
- The Stop Solution is acidic. Do not allow to contact skin or eyes. Calibrators, controls and specimen samples should be assayed in duplicate. Once the procedure has been started, all steps should be completed without interruption.
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Storage
- 4 °C,-20 °C
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Storage Comment
- -20°C. Bring all reagents to room temperature before beginning test. The kit may be stored at 4°C for immediate use within two days upon arrival. Reseal any unused strips with desiccant pack. Minimize freeze/thaw cycles.
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Expiry Date
- 12 months
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- CH25H (Cholesterol 25-Hydroxylase (CH25H))
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Alternative Name
- Cholesterol-25-Hydroxylase
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Background
- Gene Aliases: h25OH, Cholesterol 25-monooxygenase
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Gene ID
- 12642
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UniProt
- Q9Z0F5
Target See all CH25H ELISA Kits
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