hSDF1β-induced migration of NK-92 cells was assessed. NK-92 cells were incubated in a 96-well transwell plate with increasing concentrations of hSDF1β in the bottom chamber. After 2 hours, the number of NK-92 cells that migrated to the bottom chamber of the transwell was quantified by measuring DNA content using a fluorescent dye.
The purity of recombinant hSDF1β was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hSDF1β and staining overnight with Coomassie Blue.
With carrier: Lyophilized from a 0.22 μm filtered solution of 20 mM citrate, pH 3.0 containing 100 mM NaCl and 20 μg BSA per 1 μg hSDF1β. Carrier free: Lyophilized from a 0.22 μm filtered solution of 20 mM citrate, pH 3.0 containing 100 mM NaCl.
Stable in lyophilized state at 4°C for 1 year after receipt. Sterile stock solutions reconstituted with carrier protein are stable at 4°C for 2 months and at -20°C for 6 months. Avoid repeated freeze-thaw cycles.Maintain sterility. Storage at -20°C should be in a manual defrost freezer.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hSDF1β. All lots are greater than 98% pure.
Recombinant hSDF1β has a Met on the amino terminus and has a calculated MW of 8,657. DTT-reduced protein migrates as an 8 kDa polypeptide and non-reduced protein migrates as a 12 kDa polypeptide. The expected amino-terminal MKPVS of recombinant hSDF1β was verified by amino acid sequencing.
The activity of hSDF1β was determined using an NK-92 cell migration assay. The ED50 of each lot is between 0.1-0.3 µg/ml.
Less than 0.01 ng endotoxin/1 μg hSDF1β.
Recombinant human SDF1β (hSDF1β) Lys22-Met93 (Accession #NP_000600) was produced in E. coli at Cell Signaling Technology.
SDF1, also known as CXCL12 and PBSF, is an 8 kDa member of the CXC family of chemokines (1). SDF1 is highly conserved across species and the human protein differs from mouse by only one conserved amino acid substitution (2). SDF1α and SDF1β are splice variants which are identical with regard to amino acid sequence except for an additional 4 amino acids on the carboxy terminus of SDF1β (3). Both SDF1 isoforms binds to the G protein-coupled receptor, CXCR4 (4). SDF1β is a potent lymphocyte chemoattractant (5). SDF1/CXCR4 signaling is required for B cell lymphopoiesis and myelopoiesis (6,7). While CXCR4 signaling plays a key role in breast cancer metastasis, increased SDF1 expression in breast cancer may be a prognostic indicator of overall survival (8).
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