APPLICATION
Application Note
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. A proliferation assay was conducted to detect the bioactivity of recombinant human GREM1 using A549 cells. Briefly, A549 cells were seeded into triplicate wells of 96-well plates at a density of 5000 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 450nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37ºC. Proliferation of A549 cells after incubation with GREM1 for 96h observed by inverted microscope. Cell viability was assessed by CCK-8 (Cell Counting Kit-8) assay after incubation with recombinant GREM1 for 96h that GREM1 significantly increased cell viability of A549 cells.
Observed MW
25 kDa.
PROPERTIES
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Lys26~Asp184 (NP_001178252.1)
Expression System
E. coli
Purity
> 97%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET
Synonyms
gremlin 1, DAN family BMP antagonist , C15DUPq , CKTSF1B1 , CRAC1 , CRCS4 , DAND2 , DRM , DUP15q , GREMLIN , HMPS , HMPS1 , IHG-2 , MPSH , PIG2
Background
This gene encodes a member of the BMP (bone morphogenic protein) antagonist family. Like BMPs, BMP antagonists contain cystine knots and typically form homo- and heterodimers. The CAN (cerberus and dan) subfamily of BMP antagonists, to which this gene belongs, is characterized by a C-terminal cystine knot with an eight-membered ring. The antagonistic effect of the secreted glycosylated protein encoded by this gene is likely due to its direct binding to BMP proteins. As an antagonist of BMP, this gene may play a role in regulating organogenesis, body patterning, and tissue differentiation. In mouse, this protein has been shown to relay the sonic hedgehog (SHH) signal from the polarizing region to the apical ectodermal ridge during limb bud outgrowth. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jul 2010]
Database
Research Area
DATA IMAGES
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GTX00210-pro Functional Assay Image
Cell proliferation effect of GTX00210-pro Human Gremlin 1 protein (active). Cell viability was measured by Cell Counting Kit-8 (CCK-8). (A) Unstimulated A549 cells cultured in DMEM for 96hrs. (B) A549 cells cultured in DMEM, stimulated with 1 ng/ml GREM1 for 96hrs.
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GTX00210-pro Image
SDS-PAGE analysis of GTX00210-pro Human Gremlin 1 protein (active).
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GTX00210-pro Image
WB analysis of GTX00210-pro Human Gremlin 1 protein (active).
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REFERENCE
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REVIEW
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