Multiplex Assay Kit for Orexin A (OXA) ,etc. by FLIA (Flow Luminescence Immunoassay)

Hypocretin-1

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Orexin A (OXA) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Orexin A (OXA) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Orexin A (OXA) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Orexin A (OXA) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-91 88
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 79-97 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Orexin A (OXA) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 FLIA (Flow Luminescence Immunoassay) 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 FLIA (Flow Luminescence Immunoassay) 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) 98-105% 87-101% 85-93% 85-101%
EDTA plasma(n=5) 80-105% 93-105% 80-104% 85-93%
heparin plasma(n=5) 82-97% 78-93% 88-102% 89-97%

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. Preparation of standards, reagents and samples before the experiment;
2. Add 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Frontiers in Neurology Neuro-Glial and Systemic Mechanisms of Pathological Responses in Rat Models of Primary Blast Overpressure Compared to “Composite” Blast PubMed: PMC3275793
Neurological Sciences Pentylenetetrazol-induced seizures are exacerbated by sleep deprivation through orexin receptor-mediated hippocampal cell proliferation Pubmed: 23852314
Peptides Orexin-A-induced ERK1/2 activation reverses impaired spatial learning and memory in pentylenetetrazol-kindled rats via OX1R-mediated hippocampal neurogenesis. Pubmed: 24321199
Alcohol Alcohol Orexin in Patients with Alcohol Dependence Treated for Relapse Prevention: A Pilot Study PubMed: 26597795
Indian Journal of Medical Research and Pharmaceutical Sciences Evaluation of Antidiabetic And Insulintropic Potential of Nigella Sativa Seeds Water Extract and /or Alpha Lipoic Acid in Type 2 Diabetic Rats Issues%20Pdf: Vol.2
J Clin Res Pediatr Endocninol Correlation of Brain Neuropeptide (Nesfatin-1 and Orexin-A) Concentrations with Anthropometric and Biochemical Parameters in Malnourished Children PubMed: 26831553
Neural Regen Res Resuscitation therapy for traumatic brain injury-induced coma in rats: mechanisms of median nerve electrical stimulation PubMed: 26170820
Mol Med Rep Wake-promoting actions of median nerve stimulation in TBI-induced coma: An investigation of orexin-A and orexin receptor 1 in the hypothalamic region PubMed: 26059340
Mol Neurobiol Application of the Co-culture Membrane System Pointed to a Protective Role of Catestatin on Hippocampal Plus Hypothalamic Neurons Exposed to Oxygen and Glucose Deprivation article:10.1007
Neural Regeneration Research 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
Journal of Translational Medicine Recurrent circadian fasting (RCF) improves blood pressure, biomarkers of cardiometabolic risk and regulates inflammation in men Pubmed: 31426866
iraqi journal of science Relationship Between Orexin-A and Insulin Resistance in Patients with Type 2 Diabetes Mellitus
Brain Sci Changes in the Orexin System in Rats Exhibiting Learned Helplessness Behaviors 34942932
Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
RPA607Ra01 Recombinant Orexin A (OXA) Positive Control; Immunogen; SDS-PAGE; WB.
PAA607Ra01 Polyclonal Antibody to Orexin A (OXA) WB; IHC; ICC; IP.
CEA607Ra ELISA Kit for Orexin A (OXA) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA607Ra Multiplex Assay Kit for Orexin A (OXA) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.