Detection of Mouse and Rat SCF/c‑kit Ligand by Western Blot.
Western blot shows lysates of rat adipose tissue, rat liver tissue, and mouse adipose tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Mouse/Rat SCF/c‑kit Ligand Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6998) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for SCF/c‑kit Ligand at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
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: SCF/c-kit Ligand
SCF (Stem Cell Factor; also kit ligand and MGF) is a 40-45 kDa member of the SCF family of molecules. It is expressed by multiple cell types, including Sertoli cells, neurons, select fibroblasts, mast cells, melanocytes and dendritic cells. SCF binds to SCFR/c-kit (for kitten; the animal used to isolate it), and occurs in both a soluble and membrane-bound form. The soluble form of SCF produces a transient activation of SCFR, while the membrane-bound form generates a long-lasting signal in the SCFR due to the ligation and retention of SCFR at the expressing cell surface. Mature rat SCF is a 248 amino acid (aa) type I transmembrane (TM) glycoprotein (aa 26-273). It contains a 189 aa extracellular region (aa 26-214) plus a 36 aa cytoplasmic domain (aa 238-273). Proteolytic cleavage after Ala190 generates a 25-38 kDa soluble form that circulates as both a monomer and noncovalent homodimer. There is also possible cleavage after Phe183 that will create a similarly-sized isoform. An alternative splice form also exists that contains a Gly substitution for aa 174-292. This 28-32 kDa TM form may also undergo cleavage to generate a soluble 19-21 kDa isoform. Over aa 26-189, rat SCF shares 94%, 77% and 79% aa sequence identity with mouse, feline and human SCF, respectively.
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