Details
ACTIVITY TEST
Buffer Formulation 20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose and Proclin300. Traits Freeze-dried powder Purity > 97% Isoelectric Point 9.6 Applications Cell culture; Activity Assays.
Figure. Cell proliferation of A549 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. 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 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, 10µL 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 37°C. Proliferation of A549 cells after incubation with GREM1 for 96h observed by inverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8 (Cell Counting Kit-8) assay after incubation with recombinant GREM1 for 96h. The result was shown in Figure 2. It was obvious that GREM1 significantly increased cell viability of A549 cells.
(A) A549 cells cultured in DMEM, stimulated with 1ng/mL GREM1 for 96h;
(B) Unstimulated A549 cells cultured in DMEM for 96h.
Figure. Cell proliferation of A549 cells after stimulated with GREM1.
USAGE
Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) 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.
Image
Figure. Gene Sequencing (Extract)
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Figure. SDS-PAGE
Figure. Western Blot
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