Active Gremlin 1 (GREM1)

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

ACTIVITY TEST

Figure 1. Cell proliferation of MCF-7 cells after stimulated with GREM1.
Gremlin(GREM1) is an inhibitor in the TGF beta signaling pathway. GREM1and other BMP antagonists are important in the survival of cancer stroma survival and proliferation in some cancers. The protein expression is found in many cancers and is thought to play important roles in uterine cervix, lung, ovary, kidney, breast, colon, pancreas, and sarcoma carcinomas. A proliferation assay was conducted to detect the bioactivity of recombinant rat GREM1 using A549 cells. Briefly, A549 cells were seeded into triplicate wells of 96-well plates at a density of 5,000 cells/well and allowed to attach, replaced with serum-free overnight, then the medium was replaced with 2% serum standard DMEM  prior to the addition of various concentrations of GREM1. After incubated for 96h, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8(CCK-8). Briefly,10 µl of CCK-8 solution was added to each well of the plate, then the absorbance at 450 nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37 °C. Proliferation of A549 cells after incubation with GREM1 for 96h observed by inverted microscope was shown in Figure1. Cell viability was assessed by CCK-8(Cell Counting Kit-8 ) assay after incubation with recombinant GREM1 for 96h. The result was shown in Figure2. It was obvious that GREM1 significantly increased cell viability of A549 cells.(A)MCF-7 cells cultured in DMEM, stimulated with 0.001ng/ml GREM1 for 96h; (B)Unstimulated MCF7 cells cultured in DMEM for 96h.

Figure. Cell proliferation of MCF-7 cells after stimulated with GREM1.

Gremlin (GREM1) is an inhibitor in the TGF beta signaling pathway. GREM1 and other BMP antagonists are important in the survival of cancer stroma survival and proliferation in some cancers. The protein expression is found in many cancers and is thought to play important roles in uterine cervix, lung, ovary, kidney, breast, colon, pancreas, and sarcoma carcinomas. It is reported that some BMP target genes is antagonised by GREM1 and GREM2, thus a binding ELISA assay was conducted to detect the interaction of recombinant rat GREM1 and recombinant human BMP2. Briefly, GREM1 was diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 µl were then transferred to BMP2-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-GREM1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450 nm immediately. The binding activity of GREM1 and BMP2 was shown in Figure 3, the EC50 for this effect is 1.042 ug/mL.

USAGE

Reconstitute in ddH2O to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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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!)
APC128Ra01 Active Gremlin 1 (GREM1) Cell culture; Activity Assays.
RPC128Ra01 Recombinant Gremlin 1 (GREM1) Positive Control; Immunogen; SDS-PAGE; WB.
PAC128Ra01 Polyclonal Antibody to Gremlin 1 (GREM1) WB; IHC; ICC; IP.
SEC128Ra ELISA Kit for Gremlin 1 (GREM1) Enzyme-linked immunosorbent assay for Antigen Detection.
LMC128Ra Multiplex Assay Kit for Gremlin 1 (GREM1) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.
PGC128Ra01 Primer Pair for Gremlin 1 (GREM1) PCR