Multiplex Assay Kit for Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) 
Hypocretin-1
Overview
Properties
- Product No.CMA607Hu
- Organism SpeciesHomo sapiens (Human) Same name, Different species.
- ApplicationsFLIA Kit for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategoryNeuro scienceGastroenterologyHormone metabolism
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Packages (Simulation)
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Packages (Simulation)
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ISO9001: 2008, ISO13485: 2003 Registered
Recovery
Matrices listed below were spiked with certain level of recombinant Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 93-103 | 97 |
| EDTA plasma(n=5) | 93-101 | 97 |
| heparin plasma(n=5) | 84-101 | 87 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Sample | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 94-104% | 93-104% | 78-97% | 85-102% |
| EDTA plasma(n=5) | 89-96% | 89-96% | 83-101% | 90-97% |
| heparin plasma(n=5) | 97-104% | 82-97% | 78-101% | 80-101% |
Stability
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Reagents and materials provided
| Reagents | Quantity | Reagents | Quantity |
| 96-well plate | 1 | Plate sealer for 96 wells | 4 |
| Pre-Mixed Standard | 2 | Standard Diluent | 1×20mL |
| Pre-Mixed Magnetic beads (22#:OXA) | 1 | Analysis buffer | 1×20mL |
| Pre-Mixed Detection Reagent A | 1×120μL | Assay Diluent A | 1×12mL |
| Detection Reagent B (PE-SA) | 1×120μL | Assay Diluent B | 1×12mL |
| Sheath Fluid | 1×10mL | Wash Buffer (30 × concentrate) | 1×20mL |
| Instruction manual | 1 |
Assay procedure summary
1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 50μL standard or sample to each well, add 50μL prepared Detection Reagent A, add 10μL magnetic beads, and incubate 90 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
5. Wash plate once on magnetic frame;
6. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.
Test principle
Anti-Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) antibody was coated on the surface of magnetic beads. A competitive inhibition reaction is launched between biotin labeled Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) and unlabeled Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) (Standards or samples) with the pre-coated antibody specific to Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array). PE-labeled avidin was added. The amount of bound PE conjugate is reverse proportional to the concentration of Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) in the sample, the intensity of MFI developed is reverse proportional to the concentration of Orexin A (OXA) ,etc. by CBA (Cytometric Bead Array) in the sample.
Giveaways
Increment services
Citations
- Neuro-Glial and Systemic Mechanisms of Pathological Responses in Rat Models of Primary Blast Overpressure Compared to “Composite” BlastPubMed: PMC3275793
- Pentylenetetrazol-induced seizures are exacerbated by sleep deprivation through orexin receptor-mediated hippocampal cell proliferationPubmed: 23852314
- Orexin-A-induced ERK1/2 activation reverses impaired spatial learning and memory in pentylenetetrazol-kindled rats via OX1R-mediated hippocampal neurogenesis.Pubmed: 24321199
- Orexin in Patients with Alcohol Dependence Treated for Relapse Prevention: A Pilot StudyPubMed: 26597795
- Evaluation of Antidiabetic And Insulintropic Potential of Nigella Sativa Seeds Water Extract and /or Alpha Lipoic Acid in Type 2 Diabetic RatsIssues%20Pdf: Vol.2
- Correlation of Brain Neuropeptide (Nesfatin-1 and Orexin-A) Concentrations with Anthropometric and Biochemical Parameters in Malnourished ChildrenPubMed: 26831553
- Resuscitation therapy for traumatic brain injury-induced coma in rats: mechanisms of median nerve electrical stimulationPubMed: 26170820
- Wake-promoting actions of median nerve stimulation in TBI-induced coma: An investigation of orexin-A and orexin receptor 1 in the hypothalamic regionPubMed: 26059340
- Application of the Co-culture Membrane System Pointed to a Protective Role of Catestatin on Hippocampal Plus Hypothalamic Neurons Exposed to Oxygen and Glucose Deprivationarticle:10.1007
- Wake-promoting effects of vagus nerve stimulation after traumatic brain injury: upregulation of orexin-A and orexin receptor type 1 expression in the prefrontal …Pubmed:29557373
- Recurrent circadian fasting (RCF) improves blood pressure, biomarkers of cardiometabolic risk and regulates inflammation in menPubmed: 31426866
- Relationship Between Orexin-A and Insulin Resistance in Patients with Type 2 Diabetes Mellitus
- Changes in the Orexin System in Rats Exhibiting Learned Helplessness Behaviors34942932