Description
Accession
Q9QZD1
Source
Optimized DNA sequence encoding Rat Stromal Cell-Derived Factor-1 alpha mature chain was expressed in Escherichia Coli.
Molecular weight
NativeRat SDF-1 alpha, generated by the proteolytic removal of the signal peptide and propeptide,the molecule has a calculated molecular mass of approximately kDa. Recombinant Rat SDF-1 alpha is a monomer protein consisting of69 amino acid residue subunits, migrates as an approximately kDa protein under non-reducing and reducing conditions in SDS-PAGE.
Purity
>97%, as determined by SDS-PAGE and HPLC
Biological Activity
Determined by its ability to chemoattract human monocytes using a concentration range of-80 ng/ml.
Protein Sequence
MDAKVVAVLA LVLAALCISD GKPVSLSYRC PCRFFESHVA RANVKHLKIL NTPNCALQIV ARLKSNNRQV CIDPKLKWIQ EYLDKALNK
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Presentation
Rat SDF-1 alpha was lyophilized from a 0.2 μm filtered PBS solution pH7.5.
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers.
Storage
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at2° -8° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Methods
Elevated expression of CXCR4 on BM neutrophils from AnxA1−/− mice.
- C) Chemotatic response to various concentrations of CXCL12 was assessed for purified BM neutrophil populations from both WT and AnxA1−/− animals.
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Chemotaxis assay: Directed cell migration potential/chemotaxis towards SDF-1 gradient was analyzedin vitro as described [. - Briefly, 0.5x106 cells were induced by addition of 1μg/ml doxycyline in IMDM growth media and incubated for 48 hours.
- 600 μl IMDM (supplemented with 10% FBS) containing 125 ng/ml SDF-1α was added to the lower chamber of a Costar 24-wells transwell (pore size 5 μm, ).
- Then, 0.1x106 doxycycline induced cells in 100 μl medium'>IMDM medium (supplemented with 0.1% FBS) were loaded to the upper chamber and allowed to migrate for 4 hours at 37°C in a humidified CO2 incubator.
- After incubation, migrated cells were collected from lower chamber and counted on a hemocytometer under microscope.
