Multiplex Assay Kit for Corticosteroid Binding Globulin (CBG) ,etc. by CBA (Cytometric Bead Array)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 96-105 101
EDTA plasma(n=5) 79-90 79
heparin plasma(n=5) 80-93 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Corticosteroid Binding Globulin (CBG) ,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 Corticosteroid Binding Globulin (CBG) ,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 Corticosteroid Binding Globulin (CBG) ,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) 96-104% 81-98% 86-102% 97-105%
EDTA plasma(n=5) 91-105% 91-99% 84-98% 88-97%
heparin plasma(n=5) 95-102% 84-93% 98-105% 87-104%

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

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 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 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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