ELISA Kit for Spondin 2 (SPON2)

DIL1; Mindin; M-spondin; Extracellular Matrix Protein; Differentially expressed in cancerous and non-cancerous lung cells 1

Specificity

This assay has high sensitivity and excellent specificity for detection of Spondin 2 (SPON2).
No significant cross-reactivity or interference between Spondin 2 (SPON2) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Spondin 2 (SPON2) and the recovery rates were calculated by comparing the measured value to the expected amount of Spondin 2 (SPON2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 97-105 101
EDTA plasma(n=5) 96-103 101
heparin plasma(n=5) 89-97 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Spondin 2 (SPON2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Spondin 2 (SPON2) 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 Spondin 2 (SPON2) 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) 89-97% 90-99% 79-101% 87-103%
EDTA plasma(n=5) 94-102% 96-103% 99-105% 92-101%
heparin plasma(n=5) 78-95% 82-97% 81-92% 85-97%

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
The Journal of Urology Spondin-2, a secreted extracellular matrix protein, is a novel diagnostic biomarker for prostate cancer Pubmed: 23665271
Oxford Journals SP136EVALUATION OF SERUM SPONDIN 2 LEVELS IN THE DIFFERENT ETIOLOGIES OF GLOMERULAR DISEASES Content: 30
Nephrology (Carlton) Evaluation of Serum Spondin 2 Levels in the Different Stages of Type 2 Diabetic Nephropathy PubMed: 25973958
Oncogene The extracellular matrix protein mindin attenuates colon cancer progression by blocking angiogenesis via Egr-1-mediated regulation pubmed:28991232
Endocrinology Study of human T21 placenta suggests a potential role of mesenchymal spondin-2 in placental vascular development Pubmed: 30715257
Journal of Cellular and Molecular Medicine Mindin serves as a tumour suppressor gene during colon cancer progression through MAPK/ERK signalling pathway in mice Pubmed: 32614521
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
RPF396Mu01 Recombinant Spondin 2 (SPON2) Positive Control; Immunogen; SDS-PAGE; WB.
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