ELISA Kit for Gremlin 1 (GREM1) Homo sapiens (Human) Sandwich ELISA

PIG2; CKTSF1B1; DAND2; IHG-2; DAND2; DRM; Increased in high glucose protein 2; Cell proliferation-inducing gene 2 protein; Cysteine knot superfamily 1, BMP antagonist 1

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  • ELISA Kit for Gremlin 1 (GREM1) Packages (Simulation)
  • ELISA Kit for Gremlin 1 (GREM1) Packages (Simulation)
  • ELISA Kit for Gremlin 1 (GREM1) Results demonstration
  • SEC128Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Gremlin 1 (GREM1) and the recovery rates were calculated by comparing the measured value to the expected amount of Gremlin 1 (GREM1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 89-97 93
EDTA plasma(n=5) 80-89 85
heparin plasma(n=5) 78-97 83

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Gremlin 1 (GREM1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Gremlin 1 (GREM1) 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 Gremlin 1 (GREM1) 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) 97-105% 78-102% 80-96% 81-95%
EDTA plasma(n=5) 95-103% 79-90% 96-105% 80-97%
heparin plasma(n=5) 84-97% 85-101% 96-104% 91-101%

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.

ELISA Kit for Gremlin 1 (GREM1)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Gremlin 1 (GREM1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Gremlin 1 (GREM1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Gremlin 1 (GREM1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Gremlin 1 (GREM1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Intrinsic BMP Antagonist Gremlin-1 as a Novel Circulating Marker in Pulmonary Arterial HypertensionPubMed: 25926293
  • Overexpression of Gremlin promotes non-small cell lung cancer progressionPubMed: 26392110
  • Triple head-to-head comparison of fibrotic biomarkers galectin-3, osteopontin and gremlin-1 for long-term prognosis in suspected and proven acute heart failure patientsPubMed: 26539964
  • Triple head-to-head comparison of fibrotic biomarkers galectin-3, osteopontin and gremlin-1 for long-termprognosis in suspected and proven acute heart failure patientsPubmed:26539964
  • BMPR2 Mutation-independent Mechanisms of Disrupted BMP Signaling in IPAHPubmed:27187737
  • GREM1 Is a Key ReCavia (Guinea pig )lator of Synoviocyte Hyperplasia and InvasivenessPubmed:26834210
  • The BMP2 variant L51P restores the osteogenic differentiation of human mesenchymal stromal cells in the presence of intervertebral disc cells.pubmed:28266688
  • Genomic reprograming analysis of the Mesothelial to Mesenchymal Transition identifies biomarkers in peritoneal dialysis patients.pubmed:28327551
  • Bone Morphogenic Protein Type 2 Receptor Mutation-Independent Mechanisms of Disrupted Bone Morphogenetic Protein Signaling in Idiopathic Pulmonary Arterial …rcmb.2015-0402OC
  • Gremlin-1 suppression increases BMP-2-induced osteogenesis of human mesenchymal stem cellspubmed:28260028
  • Platelets as a novel source of Gremlin-1: Implications for thrombo inflammationpubmed:27929199
  • Gremlin, A Potential Urinary Biomarker of Anca-Associated Crescentic GlomerulonephritisPubmed: 31053735
  • Transcriptomic and Proteomic Analyses Reveal the Potential Mode of Action of Chondrocyte Sheets in Hyaline Cartilage RegenerationPubmed: 31878307
  • Suppression of microRNA-23a/b-3p delays the development degenerative joint disease by activating Grem1 and TGF-β/Smad signaling pathwayPubmed: 32186934
  • Protective role of microRNA-23a/b-3p inhibition against osteoarthritis through Gremlin1-depenent activation of TGF-β/smad signaling in chondrocytesaPubmed:35441352
  • CAF‐derived exosomes transmitted Gremlin‐1 promotes cancer progression and decreases the sensitivity of hepatoma cells to sorafenibPubmed:35638711

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