Products/Additional Cell Culture Products
Recombinant FGF2 1mg
Expression system/Source: Protein, purified, expressed in E.Coli, animal-free product.
Target formulation: Lyophilized from a 0.2 μM filtered solution in 50 mM Sodium Phosphate buffer, 100 mM NaCl, pH 6,0
Purity: >95%, by SDS-PAGE with Coomassie staining and HPLC
Endotoxin Level: <0.01 EU per 1 μg of the protein by the LAL method.
UniProt Accession #: P09038
Structure / Isoform: Single monomer
Predicted Molecular Mass/ Molecular weight: 19.547 kDa, determined by high-resolution TOF-MS to confirm correct protein sequence.
18 kDa, reducing condition, 18 kDa, non-reducing conditions.
UniProt Description of FGF:
Acts as a ligand for FGFR1, FGFR2, FGFR3 and FGFR4 (PubMed:8663044). Also acts as an integrin ligand which is required for FGF2 signaling (PubMed:28302677). Binds to integrin ITGAV: ITGB3 (PubMed:28302677). Plays an important role in the regulation of cell survival, cell division, cell differentiation and cell migration (PubMed:8663044, PubMed:28302677). Functions as a potent mitogen in vitro (PubMed:1721615, PubMed:3964259, PubMed:3732516). Can induce angiogenesis (PubMed:23469107, PubMed:28302677). Mediates phosphorylation of ERK1/2 and thereby promotes retinal lens fiber differentiation (PubMed:29501879).
MGSSHHHHHH SSGLVPRGSH MMAAGSITTL PALPEDGGSG AFPPGHFKDP KRLYCKNGGF FLRIHPDGRV DGVREKSDPH IKLQLQAEER GVVSIKGVCA NRYLAMKEDG RLLASKCVTD ECFFFERLES NNYNTYRSRK YTSWYVALKR TGQYKLGSKT GPGQKAILFL PMSAKS
BIOACTIVITY ASSAY SUMMARY PROTOCOL
1. Fractions of Cellaria FGF are prepared with sterile dPBS, confirming final concentrations using a NanoDrop or equivalent piece of equipment.
2. Further serial dilutions are prepared in DMEM.
3. Seed 10,000 cells per well of a 96-well plate.
4. After 24 hours, wash cells with 200uL of DMEM(Serum-Free)
5. Aspirate and add 50uL of serum free media to each well.
6. Incubate plate for 2 hours, then add 50uL of 2x FGF Titration curves to appropriate wells.
7. Incubate for 24 hours, aspirate, and add 100uL of 1x FGF Titration curves to appropriate wells.
8. Incubate for 24 hours, equilibrate for 30 minutes, then add 100uL CellTiterGlo 2.0.
9. The contents are mixed on an orbital shaker for 2 minutes, then incubated at room temperature for 10 minutes.
10. The plate can now be read by any plate reader capable of read
Full-Length Protocol Can Be Found Here: PDF
PREPARATION AND STORAGE
1. Reconstitute at 1 mg/mL in distilled water (final buffer composition 50 mM Sodium Phosphate buffer, 100 mM NaCl, pH 6,0)
2. The product is shipped with cold packs. Upon receipt, store it immediately at the temperature recommended below.
3. Avoid repeated freeze‐thaw cycles. A minimum of 6 months when stored at ≤ ‐20 °C as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, ≤ ‐20 °C under sterile conditions after reconstitution.
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