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Recombinant Human Stem Cell Factor (SCF)
(Cat. No.: C034)
Background:
Stem Cell Factor (SCF) is a hematopoietic growth factor that exerts its activity at the early stages of hematopoiesis. SCF stimulates the proliferation of myeloid, erythroid and lymphoid progenitors in bone marrow cultures and has been shown to act synergistically with colony stimulating factors.
Description:
Recombinant Human SCF produced in E. coli is a single, non-glycosylated polypeptide chain containing 165 amino acids and having a molecular mass of 18409 Dalton.
Quality Control:
Biological activity: The ED50 as determined by the dose-dependant stimulation of Human TF-1 cells is less then 2 ng/ml, corresponding to a Specific Activity of 5.0 x 105 IU/mg.
Purity: Greater than 95% as determined by
(a) Analysis by SEC-HPLC.
(b) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.
Amino-Acid Sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Met-Glu-Gly-Ile-Cys.
Endotoxin: Less than 0.1 ng/µg (IEU/µg) of rHuSCF.
Formulation: rHuSCF was lyophilized from a concentrated (1mg/ml) sterile solution containing 10mM Acetic acid.
Storage: Lyophilized rHuSCF although stable at room temperature for 3 weeks, should be stored desiccated below -18oC. Upon reconstitution rHuSCF should be stored at 4oC between 2-7 days and for future use below -18oC. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.
Reconstitution: It is recommended to reconstitute the lyophilized rHuSCF in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Latest Publications:
1: Am J Pathol 2009 Jan;
Role of Stem Cell Factor and Bone Marrow-Derived Fibroblasts in Airway Remodeling.
[Abstract] Recent evidence suggests that bone marrow-derived fibroblasts are involved in airway remodeling in asthma, but the role and mechanism of recruitment of these fibroblasts remains unclear. Stem cell factor (SCF), a key factor in the propagation of hematopoietic stem cells, is important in the process of airway remodeling as well. To test the hypothesis that SCF is involved in the recruitment and differentiation of bone marrow-derived progenitor cells, GFP-bone marrow chimeric mice were created. These mice were then sensitized and chronically challenged with cockroach antigen to induce chronic airway disease. Fluorescence microscopy revealed an influx of significant numbers of GFP-expressing fibroblasts in the airways of these mice, which was confirmed by flow
cytometric analysis of cells co-expressing both GFP and collagen I. These cells preferentially expressed c-kit, interleukin-31 receptor, and telomerase reverse transcriptase when compared with control lung-derived fibroblasts. Interestingly, SCF stimulated interleukin-31 receptor expression in bone marrow cells, whereas interleukin-31 strongly induced telomerase reverse transcriptase expression in fibroblasts. Treatment with neutralizing antibodies to SCF significantly reduced airway remodeling and suppressed the recruitment of these bone marrow-derived cells to the lung. Thus SCF in conjunction with interleukin-31 may play a significant role in airway remodeling by promoting the recruitment of bone marrow-derived fibroblast precursors into the lung with the capacity to promote lung
myofibroblast differentiation.
2: Dev Neurosci 2009 Jan;
Essential Role of Stem Cell Factor Signaling in Primary Sensory Neuron Development.
[Abstract] Here we show that stem cell factor (SCF) signaling through its receptor, c-kit, is essential for the development of c-kit-expressing small- and medium-diameter primary sensory neurons. We used the W mouse, which is c-kit deficient and has a perinatal lethal phenotype due to a naturally occurring point mutation in the c-kit gene. In c-kit-null newborn mice, 52.5% of substance P immunoreactive and 31.4% of calcitonin gene-related peptide (CGRP) immunoreactive small- and medium-diameter sensory neurons were absent, whereas large-diameter sensory neurons were unaffected. Equivalent deficits occurred during embryogenesis. There was neither a developmental delay nor degeneration of differentiated neurons. We thus conclude that, in the absence of SCF signaling,
neural crest-derived progenitors do not differentiate into c-kit-expressing visceral and somatic afferent neurons.
3: Clin Cancer Res 2009
Jan;Vol 15(1)
Overexpression of the KIT/SCF in uveal melanoma does not translate into clinical efficacy of imatinib mesylate.
[Abstract] PURPOSE: Recently, gene amplification and overexpression of KIT as well as activating mutations in the KIT gene have been described to occur in certain subsets of melanoma. These findings suggest KIT as a potential target for therapy with imatinib mesylate in these melanomas. To date, data on the KIT status in uveal melanoma (UM) is limited. EXPERIMENTAL DESIGN: We analyzed the expression of the KIT protein (CD117, c-kit) and its ligand, stem cell factor (SCF), in primary and metastatic UM. RESULTS: By immunohistochemistry, SCF-positive tumor cells (>90%) were detectable in 43% of primary UM and in 58% of UM metastases. Strong expression of KIT (>90%) in tumor cells was present in 55% of primary UM and in 76% of UM metastases. This overexpression of both KIT
and SCF suggests the clinical application of imatinib mesylate in metastatic UM. This notion was tested in a clinical study using Simon's two-stage design. Patients received imatinib (600 mg p.o. daily) until progress or unacceptable toxicities. The trial did not enter stage II as no objective response was observed in the first group. This observation prompted further molecular analysis, which revealed no mutations in the genomic sequence of KIT in exons 11, 13, 17, and 18. Moreover, the mitogen-activated protein kinase pathway was not activated in any of the tumors as measured by ERK phosphorylation. CONCLUSIONS: These results show the lack of clinical effectiveness of imatinib in UM, which was originally anticipated based on the high levels of KIT and SCF expression.
4: Genetics 2008 Dec;
Isolation and Characterization of cul1-7, a Recessive Allele of CULLIN1 that Disrupts SCF Function at the C-terminus of CUL1 in Arabidopsis thaliana.
[Abstract] Many aspects of plant biology depend on the ubiquitin proteasome system for degradation of regulatory proteins. Ubiquitin E3 ligases confer substrate specificity in this pathway, and SCF-type ligases comprise a major class of E3s. SCF ligases have four subunits: SKP1, CUL1, RBX1, and an F-box protein for substrate recognition. The Aux/IAAs are a well-characterized family of SCF substrates in plants. Here, we report characterization of a mutant isolated from a genetic screen in Arabidopsis thaliana designed to identify plants defective in degradation of an Aux/IAA fusion protein, Aux/IAA1-luciferase (IAA1-LUC). This mutant exhibited four-fold slower IAA1-LUC degradation compared to the progenitor line, and seedlings displayed altered auxin responses.
Experiments identified the mutant as an allele of CUL1, called cul1-7. The cul1-7 mutation affects the C-terminus of the protein, results in reduced cul1-7 levels, and interferes with RBX1 interaction. cul1-7 seedlings are defective in degradation of an endogenous SCF substrate, Repressor of ga1-3 (RGA), and have altered responses to gibberellins. cul1-7 seedlings have slower degradation of the light-labile red/far-red photoreceptor phytochrome A and are photomorphogenic in the dark. This mutation represents the first reported allele of CUL1 to directly affect subunit interactions at the CUL1 C-terminus.
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