Mouse C4.4A/LYPD3 Antibody

  
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse C4.4A in direct ELISAs and Western blots. In direct ELISAs, approximately 40% cross-reactivity with recombinant human C4.4A is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse C4.4A
    Leu33-His287
    Accession # Q91YK8
  • 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
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunohistochemistry
C4.4A/LYPD3 in Mouse Ovary. C4.4A/LYPD3 was detected in perfusion fixed frozen sections of mouse ovary using Goat Anti-Mouse C4.4A/LYPD3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5567) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Detection of Mouse C4.4A/LYPD3 by Western Blot. Western blot shows lysates of mouse skin tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse C4.4A/LYPD3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5567) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for C4.4A/LYPD3 at approximately 95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 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: C4.4A/LYPD3

C4.4A, also known as Ly6/PLAUR domain containing 3 (LYPD-3), is a GPI-linked protein with structural similarity to the urokinase-type plasminogen activator receptor (uPAR) (1). Mature mouse C4.4A contains two uPAR/Ly6 domains and a Ser/Thr/Pro-rich (STP) region that includes a protease sensitive site (2, 3). Mouse C4.4A shares 80% and 92% amino acid sequence identity with human and rat C4.4A, respectively. It is a 65-100 kDa molecule with cell type-specific N- and O-linked glycosylation (4, 5). Proteolytic cleavage following the second uPAR/Ly6 domain generates a 35-40 kDa soluble form, while ADAM10 or ADAM17-mediated cleavage within the STP region generates a 90 kDa soluble form (6-8). Soluble C4.4A can also be shed and released in membrane vesicles (5). C4.4A is expressed in the suprabasal layers of stratified squamous epithelium and is upregulated on migrating keratinocytes during wound healing (6, 7). Its expression is downregulated during the onset of epithelial dysplasia but subsequently upregulated at the invasive front of melanomas and various carcinomas (2, 6, 5, 9). Metastases derived from these tumors also express high levels of C4.4A (2, 5, 6). The interaction of C4.4A with Laminin-1 and -5 on neighboring cells promotes the adhesion, spreading, and migration of tumor cells (4, 6, 10). C4.4A additionally interacts with Galectin-3 and the anterior gradient proteins AG-2 and AG-3 (10, 11). C4.4A over-expression in non-small cell lung cancer is predictive of increased mortality (12).

  • References:
    1. Jacobsen, B. and M. Ploug (2008) Curr. Med. Chem. 15:2559.
    2. Smith, B.A. et al. (2001) Cancer Res. 61:1678.
    3. Wurfel, J. et al. (2001) Gene 262:35.
    4. Rosel, M. et al. (1998) Oncogene 17:1989.
    5. Paret, C. et al. (2007) Br. J. Cancer 97:1146.
    6. Hansen, L.V. et al. (2008) Int. J. Cancer 122:734.
    7. Hansen, L.V. et al. (2004) Biochem. J. 380:845.
    8. Esselens, C.W. et al. (2008) Biol. Chem. 389:1075.
    9. Seiter, S. et al. (2001) J. Invest. Dermatol. 116:344.
    10. Paret, C. et al. (2005) Int. J. Cancer 115:724.
    11. Fletcher, G.C. et al. (2003) Br. J. Cancer 88:579.
    12. Hansen, L.V. et al. (2007) Lung Cancer 58:260.
  • Long Name:
    Leucine Rich Repeat and Fibronectin Type III Domain Containing 3
  • Entrez Gene IDs:
    27076 (Human); 72434 (Mouse); 60378 (Rat)
  • Alternate Names:
    C4.4A; C4.4A2310061G07Rik; GPI-anchored metastasis-associated protein C4.4A homolog; GPI-anchored metastasis-associated protein homolog; LY6/PLAUR domain containing 3; ly6/PLAUR domain-containing protein 3; LYPD3; Matrigel-induced gene C4 protein; MIG-C4
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