Multiplex Assay Kit for Gremlin 1 (GREM1) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Gremlin 1 (GREM1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Gremlin 1 (GREM1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 101
EDTA plasma(n=5) 80-93 87
heparin plasma(n=5) 78-98 83
sodium citrate plasma(n=5) 91-102 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Gremlin 1 (GREM1) ,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 Gremlin 1 (GREM1) ,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 Gremlin 1 (GREM1) ,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) 78-97% 90-102% 78-102% 78-97%
EDTA plasma(n=5) 81-91% 87-101% 94-105% 93-105%
heparin plasma(n=5) 80-99% 93-105% 85-94% 79-98%
sodium citrate plasma(n=5) 94-103% 94-102% 97-104% 82-92%

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#:GREM1) 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
Lung Intrinsic BMP Antagonist Gremlin-1 as a Novel Circulating Marker in Pulmonary Arterial Hypertension PubMed: 25926293
Tumour Biol Overexpression of Gremlin promotes non-small cell lung cancer progression PubMed: 26392110
Int J Cardiol 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 patients PubMed: 26539964
International Journal of Cardiology Triple head-to-head comparison of fibrotic biomarkers galectin-3, osteopontin and gremlin-1 for long-termprognosis in suspected and proven acute heart failure patients Pubmed:26539964
American Journal of Respiratory Cell and Molecular Biology BMPR2 Mutation-independent Mechanisms of Disrupted BMP Signaling in IPAH Pubmed:27187737
J Rheumatol GREM1 Is a Key ReCavia (Guinea pig )lator of Synoviocyte Hyperplasia and Invasiveness Pubmed:26834210
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
Sci Rep. Genomic reprograming analysis of the Mesothelial to Mesenchymal Transition identifies biomarkers in peritoneal dialysis patients. pubmed:28327551
American Journal of Respiratory Cell and Molecular Biology Bone Morphogenic Protein Type 2 Receptor Mutation-Independent Mechanisms of Disrupted Bone Morphogenetic Protein Signaling in Idiopathic Pulmonary Arterial … rcmb.2015-0402OC
Molecular Medicine Reports Gremlin-1 suppression increases BMP-2-induced osteogenesis of human mesenchymal stem cells pubmed:28260028
Cellular Haemostasis and Platelets Platelets as a novel source of Gremlin-1: Implications for thrombo inflammation pubmed:27929199
Scientific Reports Gremlin, A Potential Urinary Biomarker of Anca-Associated Crescentic Glomerulonephritis Pubmed: 31053735
International Journal of Molecular Sciences Transcriptomic and Proteomic Analyses Reveal the Potential Mode of Action of Chondrocyte Sheets in Hyaline Cartilage Regeneration Pubmed: 31878307
American Journal of Physiology-Cell Physiology Suppression of microRNA-23a/b-3p delays the development degenerative joint disease by activating Grem1 and TGF-β/Smad signaling pathway Pubmed: 32186934
Inflammopharmacology Protective role of microRNA-23a/b-3p inhibition against osteoarthritis through Gremlin1-depenent activation of TGF-β/smad signaling in chondrocytesa Pubmed:35441352
Mol Carcinog CAF‐derived exosomes transmitted Gremlin‐1 promotes cancer progression and decreases the sensitivity of hepatoma cells to sorafenib Pubmed:35638711
Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
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