Multiplex Assay Kit for Corticosteroid Binding Globulin (CBG) ,etc. by FLIA (Flow Luminescence Immunoassay)

SERPINA6; Transcortin; Serpin A6; Serpin Peptidase Inhibitor,Clade A,Member 6(Alpha-1 Antiproteinase; Antitrypsin)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 88-95 92
EDTA plasma(n=5) 82-97 89
heparin plasma(n=5) 83-93 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Corticosteroid Binding Globulin (CBG) ,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 Corticosteroid Binding Globulin (CBG) ,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 Corticosteroid Binding Globulin (CBG) ,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) 84-98% 78-94% 87-101% 78-89%
EDTA plasma(n=5) 88-101% 92-102% 98-105% 95-105%
heparin plasma(n=5) 89-96% 79-94% 96-105% 92-104%

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#:CBG) 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 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
General Endocrinology The Regulation of Rasd1 Expression by Glucocorticoids and Prolactin Controls Peripartum Maternal Insulin Secretion Endjournals: Source
PLoS One Modulation in Wistar Rats of Blood Corticosterone Compartmentation by Sex and a Cafeteria Diet PubMed: PMC3579843
Annals of Anatomy Differences in colocalization of corticosteroid-binding globulin and glucocorticoid receptor immunoreactivity in the rat brain ScienceDirect: S0940960212001628
Physiological Reports Adrenocortical control in the neonatal rat: ACTH‐and cAMP‐independent corticosterone production during hypoxia Pubmed: 24303136
Shock. Adrenocorticotropic Hormone But Not High-Density Lipoprotein Cholesterol or Salivary Cortisol Was a Predictor of Adrenal Insufficiency in Patients With Septic Shock. Pubmed: 24667620
Neurobiology of Aging Greater glucocorticoid receptor activation in hippocampus of aged rats sensitizes microglia Pubmed:25559333
Cell Death Differ. Novel p53 target genes secreted by the liver are involved in non-cell-autonomous reCavia (Guinea pig )lation Pubmed:26358154
Journal of Exercise, Sports & Orthopedics Impact of Season and Oral Contraceptive use on Cortisol Levelsin Physically Active Women exercise-sports-orthopedics:exercise-sports-orthop
Impact of Season and Oral Contraceptive use on Cortisol Levels in Physically Active Women diva2:1047030
Neuropharmacology Impaired glucocorticoid-mediated HPA axis negative feedback induced by juvenile social isolation in male rats Pubmed:29407214
Pharmacological Reports The impact of early-life stress on corticosteroid carrier protein levels and 11β-hydroxysteroid dehydrogenase 1 expression in adolescent rats
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