Detection of Mouse G‑CSF R/CD114 by Western Blot. Western blot shows lysates of ST‑2 mouse bone marrow-derived stromal cell line. PVDF Membrane was probed with 0.1 µg/mL of Sheep Anti-Mouse G‑CSF R/CD114 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6039) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for G‑CSF R/CD114 at approximately 110 and 112 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: G-CSF R/CD114
Granulocyte colony stimulating factor (G-CSF) is a pleiotropic cytokine best known for its specific effects on the proliferation, differentiation, and activation of hematopoietic cells of the neutrophilic and granulocyte lineage (1). G-CSF plays an important role in defense against infection, in inflammation and repair, and in the maintenance of steady state hematopoiesis. Cell activation by G‑CSF is mediated by granulocyte colony stimulating factor receptor alpha (G-CSF R; also CD114), a 95‑105 kDa type I transmembrane protein and member of the cytokine receptor superfamily, type I cytokine receptor family, and type 2 subfamily of receptor proteins. Mouse G‑CSF R is synthesized as an 837 amino acid (aa) precursor that contains a 25 aa signal sequence, a 601 aa extracellular domain (ECD), a 24 aa transmembrane region, and a 187 aa cytoplasmic tail. The ECD contains one Ig-like C2-type domain, five fibronectin type-III domains, and 11 potential sites for N‑linked glycosylation. Within the ECD there is also a WSXWS motif (aa 319‑323) that is necessary for proper protein folding and thereby efficient intracellular transport and cell-surface receptor binding (2). Also, within the cytoplasmic domain there is a Box 1 motif which is required for JAK interaction and/or activation (1). Mouse G‑CSF R shares 63% aa sequence identity with human G‑CSF R. G-CSF R is expressed in mature neutrophils, neutrophilic precursors, myeloid leukemia cells, and placenta (1). Mutations have been found in the gene encoding G-CSF R in some patients with severe congenital neutropenia (1). These mutations typically lead to a truncation in the cytoplasmic domain of the G-CSF R leading to maturation arrest of neutrophilic precursors in the bone marrow and neutropenia in peripheral blood (3). Binding of G-CSF to its receptor induces dimerization or oligomerization of the receptor activating cytoplasmic tyrosine kinases (2). Signal transduction from pathways that involve Janus tyrosine kinases/signal transducer and activator of transcription proteins (Jak1, Jak2, and Tyk2/STAT3 and STATG), src-related protein tyrosine kinases (Lyn and Syk), Ras/MAP kinase, and phosphatidylinositol have been reported to be activated upon G-CSF stimulation (4).
Ward, A.C. (2007) Front. Biosci. 12:608.
Layton, J.E. and N.E. Hall (2006) Front. Biosci. 11:3181.
Mitsui, T. et al. (2003) Blood 101:2990.
Nicola, N.A. in Cytokine Reference, 2001, Oppenheim, J.J. and M. Feldmann, eds. Academic Press p.1935.
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