Detects human SPRED2 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant human (rh) SPRED1 is observed and less than 3% cross-reactivity with rhSPRED4 is observed.
Polyclonal Sheep IgG
E. coli-derived recombinant human SPRED2 Lys117-Ser297 Accession # Q7Z698
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Detection of Human SPRED2 by Western Blot.
Western blot shows lysates of Hep3B human hepatocellular carcinoma cell line. PVDF Membrane was probed with 1 µg/mL of Human SPRED2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4819) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for SPRED2 at approximately 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
SPRED2 in PC‑12 Rat Cell Line.
SPRED2 was detected in immersion fixed PC‑12 rat adrenal pheochromocytoma cell line using Human SPRED2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4819) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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.
SPRED2 (Sprouty-related protein with an EVH1 domain-2) is a 47-53 kDa member of the SPRED family of regulatory molecules. In human, it is found in keratinocytes, intestinal columnar epithelium, and salivary duct epithelium. It inhibits receptor tyrosine kinase (RTK) signaling by blocking Ras activation of Raf. This is achieved by inducing the lysosomal degradation of RTKs following SPRED2 binding to NBR1. Human SPRED2 is 418 amino acids (aa) in length. It contains an N‑terminal WH1/EVH1 domain that is involved in ERK inhibition (aa 5-122), a central KBD domain that binds to the SCFR (aa 201-257), and a C-terminal Sprouty‑related domain that mediates homodimerization, and heterodimerization with SPRED1 (aa 308-416). SPRED is ubiquitinated, and undergoes phosphorylation on Ser, Thr and Tyr residues. There is one splice variant that shows a six aa substitution for aa 1-9. Over aa 117-297, human SPRED2 shares 84% aa identity with mouse SPRED2.
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