Multiplex Assay Kit for Corticotropin Releasing Factor (CRF) ,etc. by FLIA (Flow Luminescence Immunoassay) Sus scrofa; Porcine (Pig) Multiplex ELISA

CRH; Corticotropin Releasing Hormone; Corticoliberin

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

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  • Multiplex Assay Kit for Corticotropin Releasing Factor (CRF) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Corticotropin Releasing Factor (CRF) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Corticotropin Releasing Factor (CRF) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA835Po.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 93-101 96
EDTA plasma(n=5) 83-96 90
heparin plasma(n=5) 78-94 86
sodium citrate plasma(n=5) 96-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Corticotropin Releasing Factor (CRF) ,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 Corticotropin Releasing Factor (CRF) ,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 Corticotropin Releasing Factor (CRF) ,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) 88-97% 78-89% 86-96% 78-97%
EDTA plasma(n=5) 94-102% 88-101% 94-101% 86-98%
heparin plasma(n=5) 96-104% 89-99% 84-95% 91-101%
sodium citrate plasma(n=5) 93-101% 90-103% 81-93% 87-103%

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#:CRF) 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.

Multiplex Assay Kit for Corticotropin Releasing Factor (CRF) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards,Labeled antigen and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest.A competitive inhibition reaction is launched between biotin labeled analytes of interest and unlabeled analytes of interest (Standards or samples) with the pre-coated antibody specific to analytes of interest. Following a wash to remove any unbound substances, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE) is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer. The MFI developed is reverse proportional to the concentration of analytes of interest in the sample.

Giveaways

Citations

  • Regulation of corticosterone secretion is modified by PFOS exposure at different levels of the hypothalamic–pituitary–adrenal axis in adult male ratsPubmed:24440345
  • Immune dysfunction in bipolar disorder and suicide risk: is there an association between peripheral corticotropin‐releasing hormone and interleukin‐1β?Pubmed:24862833
  • Differential stress response in rats subjected to chronic mild stress is accompanied by changes in CRH-family gene expression at the pituitary levelPubmed:25236411
  • Acute Stress-Induced Changes in Follicular Dermal Papilla Cells and Mobilization of Mast Cells:Implications for Hair Growth.pubmed:27746640
  • Heat stress-induced neuroinflammation and aberration in monoamine levels in hypothalamus are associated with temperature dysregulationpubmed:28663093
  • Colonic Transit Time and Gut Peptides in Adult Patients with Slow and Normal Colonic Transit Constipationpubmed:29082242
  • Effects of n-3 PUFA enriched and n-3 PUFA deficient diets in naïve and Aβ-treated female ratsPubmed:30028991
  • Vaccinium bracteatum Leaf Extract Reverses Chronic Restraint Stress-Induced Depression-Like Behavior in Mice: Regulation of Hypothalamic-Pituitary …Pubmed:30038568
  • Perfluorooctane sulfonate (PFOS) can alter the hypothalamic-pituitary-adrenal (HPA) axis activity by modifying CRF1 and glucocorticoid receptorsPubmed:29807116
  • The depressive-like behaviors of chronic unpredictable mild stress-treated mice, ameliorated by Tibetan medicine Zuotai: involvement in the hypothalamic …Pubmed:29379286
  • Hypothalamus-pituitary-adrenal axis involves in anti-viral ability through regulation of immune response in piglets infected by highly pathogenic porcine …Pubmed:29540178
  • Bioscience Research
  • Contribution of DNA methylation in chronic stress–induced cardiac remodeling and arrhythmias in micePubmed: 31431066
  • Altered glucocorticoid metabolism represents a feature of macroph‐agingPubmed: 32463582
  • Impairment of CRH in the intestinal mucosal epithelial barrier of pregnant Bama miniature pig induced by restraint stress33408312
  • O-GlcNAc/Akt pathway regulates glucose metabolism and reduces apoptosis in liver of piglets with acute cold stress33651993

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