High Sensitive ELISA Kit for Noggin (NOG)

SYM1; SYNS1; Synostoses(Multiple)Syndrome 1; Symphalangism 1(Proximal)

Specificity

This assay has high sensitivity for detection of High Sensitive Noggin (NOG).
100% cross-reactivity of High Sensitive Noggin (NOG) was observed among Human, Mouse, Rat.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 92-103 98
EDTA plasma(n=5) 81-89 84
heparin plasma(n=5) 98-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Noggin (NOG) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Noggin (NOG) 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 High Sensitive Noggin (NOG) 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) 88-101% 84-103% 89-96% 80-99%
EDTA plasma(n=5) 91-98% 78-103% 96-104% 97-104%
heparin plasma(n=5) 80-104% 79-101% 87-104% 84-98%

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 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

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

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Magazine Citations
Arthritis Rheumatol Imbalance between BMP2 and Noggin induces abnormal osteogenic differentiation of mesenchymal stem cells in ankylosing spondylitis PubMed: 26413886
Arthritis Rheumatol.  Imbalance Between Bone Morphogenetic Protein 2 and Noggin Induces Abnormal Osteogenic Differentiation of Mesenchymal Stem Cells in Ankylosing Spondylitis Pubmed:26413886
Clin Exp Pharmacol Physiol. Oxygen-glucose deprivation preconditioning protects neurons against oxygen-glucose deprivation/reperfusion induced injury via bone morphogenetic protein-7 mediated ERK, p38 and Smad signalling pathways. Pubmed:26385023
European cells & materials The BMP2 variant L51P restores the osteogenic differentiation of human mesenchymal stromal cells in the presence of intervertebral disc cells. pubmed:28266688
Arthritis & Rheumatology mbalance Between Bone Morphogenetic Protein 2 and Noggin Induces Abnormal OsteogenicDifferentiation of Mesenchymal Stem Cells in Ankylosing Spondylitis. pubmed:26413886
Biotechnology and Applied Biochemistry The BMP antagonist Noggin is produced by osteoblasts in response to the presence of prostate cancer cells. pubmed:28981962
Heliyon Efficient in vitro delivery of Noggin siRNA enhances osteoblastogenesis pubmed:29167826
Journal of Cellular Biochemistry Low‐intensity pulsed ultrasound promotes bone morphogenic protein 9‐induced osteogenesis and suppresses inhibitory effects of inflammatory cytokines on cellular … Pubmed: 31006911
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