Detection of Mouse Ryk by Western Blot.
Western blot shows lysates of RAW 264.7 mouse monocyte/macrophage cell line, NIH‑3T3 mouse embryonic fibroblast cell line, and EL‑4 mouse lymphoblast cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Mouse Ryk Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4649) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Ryk at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Detection of Ryk in RAW 264.7 Mouse Cell Line by Flow Cytometry.
RAW 264.7 mouse monocyte/macrophage cell line was stained with Sheep Anti-Mouse Ryk Affinity-purified Polyclonal Antibody (Catalog # AF4649, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127).
Ryk in RAW 264.7 Mouse Cell Line.
Ryk was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Sheep Anti-Mouse Ryk Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4649) 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 cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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.
Ryk (related to tyrosine (Y) kinase; also VIK and MRK) is a member of the tyrosine protein kinase family. It is a type I transmembrane glycoprotein that binds Wnt and forms a Wnt receptor complex with frizzled. In this capacity, it serves as a link between Wnt and Dishevelled. Mouse Ryk is 594 amino acids (aa) in length. It contains a 177 aa extracellular region (aa 35‑211) that shows one WIF-1-like domain (aa 51‑180), and a cytoplasmic region that possesses a nonfunctional Ser/Thr protein kinase domain (aa 318‑581). There is one alternate start site at Met117. Over aa 35‑211, mouse Ryk is 94% aa identical to human Ryk, and shows absolute (100%) aa identity to rat Ryk.
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