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ELISA Kit for Relaxin (RLN)

RLXH1; Relaxin H1; Prorelaxin H1

Specificity

This assay has high sensitivity and excellent specificity for detection of Relaxin (RLN).
No significant cross-reactivity or interference between Relaxin (RLN) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Relaxin (RLN) and the recovery rates were calculated by comparing the measured value to the expected amount of Relaxin (RLN) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-101 89
EDTA plasma(n=5) 78-99 84
heparin plasma(n=5) 84-105 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Relaxin (RLN) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Relaxin (RLN) 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 Relaxin (RLN) 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) 95-103% 85-93% 81-103% 83-91%
EDTA plasma(n=5) 88-95% 79-98% 80-105% 78-101%
heparin plasma(n=5) 90-101% 94-102% 79-105% 87-94%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Reproducao & Climatério Culture of human granulosa cells with follicular phase phenotype: influence of chemically defined system on morphology, ultrastructure, secretion of steroids and relaxin ScienceDirect: S1413208713000034
BioMed Research International Upregulation of Relaxin after Experimental Subarachnoid Hemorrhage in Rabbits Pubmed:25133183
Advanced Healthcare Materials Impact of Hydrogel Elasticity and Adherence on Osteosarcoma Cells and Osteoblasts Pubmed: 30838809
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