• Species Reactivity

  Human, Mouse

• Specificity

  Detects human and mouse Frizzled-4 in Western blots. In Western blots, no cross-reactivity with recombinant mouse (rm) Frizzled-1, -2, -3, -6, -7, -8, or recombinant human Frizzled-5 is observed. Detects Frizzled-4 by flow cytometry and immunohistochemistry on mouse differentiated embryoid bodies derived from the mouse embryonic cell line, D3.    
   

• Source

  Monoclonal Rat IgG    _2A Clone # 145901

• Purification

  Protein A or G purified from hybridoma culture supernatant

• Immunogen

  Mouse myeloma cell line NS0-derived recombinant mouse Frizzled-4    
  Phe37-Glu180    
  Accession # Q61088

• Formulation

  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.

• Label

  Unconjugated

Applications

• Recommended    
  Concentration

  Sample

• Western Blot

  1 μg/mL

  Recombinant Human Frizzled-4 (Catalog # )    
  Recombinant Mouse Frizzled-4 Fc Chimera (Catalog # )

• 
  

• Flow Cytometry

  2.5 μg/10    ⁶ cells

  HEK293 human embryonic kidney cell line

• 
  

• CyTOF-ready

  Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

• 
  

• Immunocytochemistry

  8-25 μg/mL

  Immersion fixed HEK293 human embryonic kidney cell line

• 
  

Please Note: Optimal dilutions should be determined by each laboratory for each application.  are available in the Technical Information section on our website.

Preparation and Storage

• Reconstitution

  Reconstitute at 0.5 mg/mL in sterile PBS.

• Shipping

  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: Frizzled-4

Frizzled-4, designated CD344, is a 7-transmembrane glycoprotein of the Frizzled family within the G-protein coupled receptor superfamily (1, 2). Frizzled proteins function as receptors for Wnt proteins and can activate canonical Wnt/beta-catenin signaling as well as planar cell polarity and calcium flux pathways (1). Frizzled-4 is particularly important in angiogenic Wnt pathway signaling (1, 5). Frizzleds contain a divergent N-terminal signal peptide, a highly conserved ~120  amino acid (aa) cysteine-rich domain (CRD), a variable length linker region, seven transmembrane domains, and a variable-length C-terminal tail (1). The mouse Frizzled-4 cDNA encodes 537 aa with a 36 aa signal sequence and a 186 aa N-terminal extracellular sequence (4). The portion expressed includes aa 37-180, and shares 93%, 94%, 90%, 89% and 88% identity with human, rat, equine, bovine and canine Frizzled-4, respectively. This portion competes for Wnt binding with endogenous receptors. In humans, a 122 aa soluble form that diverges at aa 95 is proposed to be a positive regulator of Wnt signaling pathways (5). Frizzled-4 is unusual in binding a non-wnt ligand, Norrin, in addition to binding Wnt ligands (1, 3, 6). Norrin binds the Frizzled-4 CRD, activates Wnt signaling pathways and uses LRP5/6 as co-receptors (3, 6). Deletion of either Frizzled-4 or Norrin in mice results in a similar phenotype including malformation of vasculature in the retina, cerebellar degeneration, and loss of hair cells in the inner ear (1, 3, 7). In humans, blindness due to familial exudative vitreoretinopathy (FEVR) is associated with mutations producing loss of function of Frizzled-4 or Norrin, designated EVR1 and EVR2, respectively (1, 3, 8). Frizzled-4 expression has been found in many tissues, including mouse ovary, where it influences corpus luteum vasculogenesis and is necessary for fertility (4, 9).

• References:

1. Huang, H-C. and P.S. Klein (2004) Genome Biol. 5:234.

2. Parmalee, N.L. and J. Kitajewski (2008) Curr. Drug Targets 9:558.

3. Xu, Q. et al. (2004) Cell 116:883.

4. Wang, Y. et al. (1996) J. Biol. Chem. 271:4468.

5. Sagara, N. et al. (2001) Biochem. Biophys. Res. Commun. 282:750.

6. Smallwood, P.M. et al. (2007) J. Biol. Chem. 282:4057.

7. Wang, Y. et al. (2001) J. Neurosci. 21:4761.

8. Robitaille, J. et al. (2002) Nat. Genet. 32:326.

9. Hsieh, M. et al. (2005) Biol. Reprod. 73:1135.

• Entrez Gene IDs:

  8322 (Human); 14366 (Mouse)

• Alternate Names:

  CD344 antigen; CD344; EVR1; exudative vitreoretinopathy 1; FEVR; frizzled (Drosophila) homolog 4; frizzled homolog 4 (Drosophila); Frizzled4; Frizzled-4; Fz-4; FZD4; FZD4S; FzE4; GPCR; hFz4; MGC34390; WNT receptor frizzled-4