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Human SDF, amino acids Lys22-Lys89 (Accession # NM_199168) was expressed in E. coli.
The 68 amino acid recombinant protein has a predicted molecular mass of approximately 7963.4 Da. The DTT-reduced protein migrates at approximately 7kDa and the non-reduced protein migrates at approximately 9kDa by SDS-PAGE. The N-terminal amino acid is Lysine.
Purity is >98%, as determined by Coomassie stained SDS-PAGE.
0.22µm filtered protein solution is in 20mM Tris pH 8.0, 0.5M NaCl.
Endotoxin level is <0.1 EU/µg (<0.01ng/µg) protein as determined by the LAL method.
10-100µg sizes are bottled at 200µg/mL. 500µg sizes and larger are bottled at the concentration indicated on the vial.
Storage & Handling:
Unopened vial can be stored at 4°C for three months, at -20°C for six months, or at -70°C for one year. For maximum results, quick spin vial prior to opening. Stock solutions should be prepared at no less than 10µg/mL in buffer containing carrier protein such as 1% BSA or HSA or 10% FBS. For long term-storage, aliquot into polypropylene vials and store in a manual defrost freezer. Avoid repeated freeze/thaw cycles.
Bioactivity was measured by its property to chemoattract resting human T cells in a dose dependent manner. The ED50 is 80 - 120 ng/ml, corresponding to a specific activity of 1.25-0.83 x 104 units/mg.
Human SDF-1 belongs to the CXCL chemokine family. The mouse cDNA SDF was initially cloned from a bone marrow stromal cell library, and the human was cloned from a pro-B-cell cDNA library. SDF is expressed by many organs, and it is most abundantly expressed in pancreas, spleen, ovary and small intestine. SDF-1 (CXCL12) and its receptor CXCR4 are involved in regulation of migration, survival, and development of multiple cell types, including human hematopoietic CD34+/CD38-/low and stromal STRO-1+ stem cells. Stress-induced modulations in SDF-1 and CXCR4 levels participate in recruitment of immature and maturing leukocytes from the BM reservoir to damaged organs as part of host defense and repair mechanism. SDF-1 (CXCL12)/CXCR4 system is involved in the establishment of organ metastasis in different cancers, for example in lymph node metastasis in breast cancer and oral squamous cell carcinoma (SCC), and peritonel metastasis in ovarian cancer. Recently, several studies have demonstrated the existence of a small subset of cancer cells which share many characteristics with stem cells and named cancer stem cells (CSC). They constitute a reservoir of self-sustaining cells with the ability to maintain the tumor growth. Most of them express CXCR4 receptor and respond to a chemotactic gradient of its specific ligand SDF-1, suggesting that CSC probably represent a subpopulation capable of initiating metastasis.
CXCL12 is expressed by intestinal epithelial cells, human brain microvessel endothelial cells (HBMEC), bone marrow stromal and epithelial cells
CXCL12 is chemotactic for both peripheral blood T cells and lamina propia T cells. CXCL12 is chemotactic for peripheral blood B cells from healthy and RA donors. CXCL12 and its receptor are involved in the mobilization of CD34+ progenitors to the peripheral blood and the homing of stem cells to their specific niches. CXCL12-induced proliferation in pancreatic intraepithelial neoplasia cells (PanIN). CXCL12 is a constitutive chemokine involved in lung, brain, joint, brain, and intestine inflammation.
SDF-1 binds to the CXCR4 expressed cells such as T lymphocytes, B cells monocytes, macrophages, tissue-committed stem/progenitor cells (TCSCs), mast cells, and vascular smooth muscle cells (VSMCs)
1. Shirozu M, et al. 1995 Genomics 28:495-500. 2. Dar A, et al. 2006 Exp Hemathol 34:967-975. 3. Muller A, et al. 2001 Nature 410(6824):50-56. 4. Gelmini S, et al. 2008 J Endocrinol Invest 31:809-819. 5. Zlotnik A, et al. 2008 J Pathol 215:211-213.
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