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Multiplex Assay Kit for 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) ,etc. by FLIA (Flow Luminescence Immunoassay)

11DH; 11-beta-HSD1; HDL; HSD11; HSD11B; HSD11L; SDR26C1; Corticosteroid 11-beta-dehydrogenase isozyme 1; Short Chain Dehydrogenase/Reductase Family 26C,Member 1

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

Specificity

This assay has high sensitivity and excellent specificity for detection of 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-97 94
EDTA plasma(n=5) 80-98 92
heparin plasma(n=5) 79-103 80
sodium citrate plasma(n=5) 81-96 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) ,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 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) ,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 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) ,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) 89-96% 89-98% 85-96% 82-93%
EDTA plasma(n=5) 78-102% 79-104% 90-97% 96-103%
heparin plasma(n=5) 83-99% 79-104% 83-104% 86-103%
sodium citrate plasma(n=5) 85-96% 80-90% 98-105% 80-89%

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#:HSD11b1) 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
Journal of Endocrinology Uninephrectomy reduces 11β-hydroxysteroid dehydrogenase type 1 and type 2 concomitantly with an increase in blood pressure in rats PubMed: 22739210
Evid Based Complement Alternat Med Anti-Inflammatory Property of PubMed: 26300946
Molecules Piper sarmentosum Effects on 11β-Hydroxysteroid Dehydrogenase Type 1 Enzyme in Serumand Bone in Rat Model of Glucocorticoid-Induced Osteoporosis. pubmed:27854305
Bioorganic Chemistry A novel derivatives of thiazol-4 (5H)-one and their activity in the inhibition of 11β-hydroxysteroid dehydrogenase type 1 Pubmed:29738970
Malaysian Journal of Medical Sciences An Evidence-Based Review: The Effects of Malaysian Traditional Herbs on Osteoporotic Rat Models
2.583 Application of ELISA Technique and Human Microsomes in the Search for 11β-Hydroxysteroid Dehydrogenase Inhibitors Doi: BioMed Research International
Chemistry & Biodiversity Synthesis of the N‐methyl derivatives of 2‐aminothiazol‐4 (5H)‐one and their interactions with 11βHSD1‐molecular modeling and in vitro studies Pubmed: 31012543
Oxidative Medicine and Cellular Longevity Hepatoprotective Effects of Morinda citrifolia Leaf Extract on Ovariectomized Rats Fed with Thermoxidized Palm Oil Diet: Evidence at Histological and Ultrastructural … Pubmed: 31827717
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
RPC268Mu01 Recombinant 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) Positive Control; Immunogen; SDS-PAGE; WB.
SEC268Mu ELISA Kit for 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) Enzyme-linked immunosorbent assay for Antigen Detection.
LMC268Mu Multiplex Assay Kit for 11-Beta-Hydroxysteroid Dehydrogenase Type 1 (HSD11b1) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.