Mouse Cripto Antibody

  
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse Cripto in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant human (rh) ART, recombinant mouse (rm) ART, rmArtemin, recombinant drosophila DPP, rhGDNF, recombinant rat (rr) GDNF, rhLAP, rhLatent TGF-beta 1, rhTGF-beta 2, rhLefty-A, rmLefty-1, rrMIS, rhNeurturin, rmNodal, rhTGF-alpha, rhTGF-beta 1, rhTGF-beta 1.2, rhTGF-beta 3, or recombinant amphibian TGF-beta 5 is observed.
  • Source
    Monoclonal Rat IgG2B Clone # 237603
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse Cripto
    Arg26-Gln150
    Accession # P51865
  • 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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Flow Cytometry
    0.25 µg/106 cells
    D3 mouse embryonic stem cell line
  • Blockade of Receptor-ligand Interaction
    In a functional ELISA, 0.1-0.4 µg/mL of this antibody will block 50% of the binding of 50 ng/mL of Recombinant Mouse Cripto (Catalog # 1538-CR) to immobilized Recombinant Mouse Activin RIB/ALK-4 Fc Chimera (Catalog # 1477-AR) coated at 1 µg/mL (100 µL/well). At 3.0 μg/mL, this antibody will block >90% of the binding.
  • 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
    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
Immunocytochemistry
Cripto in D3 Mouse Cell Line. Cripto was detected in immersion fixed D3 mouse embryonic stem cell line using Rat Anti-Mouse Cripto Monoclonal Antibody (Catalog # MAB1538) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and cell secretion. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.
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: Cripto

Cripto is the founding member of the epidermal growth factor-CriptoFRL1Cryptic (EGF-CFC) family of signaling proteins that function in cancer and various developmental processes. These developmental processes include: formation of the germ layers and dorsal organizer, specification of anterior-posterior and left-right axes, and differentiation of heart muscle (1, 2). Other members of the EGF-CFC family include Cryptic, Xenopus FRL-1 and zebrafish OEP (one-eyed pinhead). Overall sequence identity between members of the family is low, but they do share several common domains: a variant EGF-like motif, a novel conserved cysteine‑rich domain (called CFC domain), and a C-terminal hydrophobic region. Most EGF-CFC members have a glycosyl-phosphatidylinositol (GPI) anchoring site at the
C-terminus and exist as extracellular membrane-anchored proteins. However, naturally-occurring soluble isoforms also exist. Mouse Cripto shares 66% and 34% amino acid identity with human Cripto and zebrafish OEP, respectively (2). Despite weak conservation in amino acid identity, EGF-CFC family members appear to function similarly in assays for phenotypic rescue of zebrafish oep mutants (2). Both secreted and membrane bound forms of Cripto demonstrate biological activity (3). Cripto, also known as CFC-2 or TDGF-1 (teratocarcinoma-derived growth factor), was originally isolated from an undifferentiated human teratocarcinoma cell line as a potential oncogene. It is overexpressed in many types of cancers and acts as a growth factor for tumors (4). Genetic evidence from mice and zebrafish points to a role for Cripto as an essential cofactor in Nodal signaling. Cripto and OEP mutants display defects in mesoderm induction and heart morphogenesis, similar to phenotypes seen in Nodal mutants (2). Cripto acts as a cofactor for Nodal by recruiting the Activin type I Receptor, ALK-4, leading to an Act RIIB‑ALK4‑Cripto‑Nodal complex for signaling (1, 3). Cripto also forms a complex with activin and Act RIIs to block activin signaling (5). Work has shown that other TGF-beta superfamily members such as Vg1 and GDF-1 also require EGF-CFC cofactors (6). Cripto can also activate mitogen-activated protein kinase (MAPK) and Akt pathways independently of Nodal by directly binding to a membrane-associated heparan sulfate proteoglycan, glypican-1 (7).

  • References:
    1. Rosa, F.M. (2002) Science’s STKE http://stke.sciencemag.org/.
    2. Shen, M. and A. Schier (2000) Trends Genet. 16:303.
    3. Yan, Y-T. et al. (2002) Mol. Cell Biol. 22:4439.
    4. Salomon, D. et al. (2000) Endocrine-Rel. Cancer 7:199.
    5. Gray, P.C. et al. (2003) Proc. Natl. Acad. Sci. USA 100:5193.
    6. Cheng, S. et al. (2003) Genes & Dev. 17:31.
    7. Bianco, C. et al. (2003) Cancer Research 63:1192.
  • Long Name:
    Teratocarcinoma-derived Growth Factor/Cripto Growth Factor
  • Entrez Gene IDs:
    6997 (Human); 21667 (Mouse)
  • Alternate Names:
    CFC-2; CR; Cripto; Cripto-1 growth factor; Cripto-1; CRIPTOCRGF; Epidermal growth factor-like cripto protein CR1; TDGF1; TDGF-1; teratocarcinoma-derived growth factor 1
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