Mouse Cripto Antibody

Catalog #: AF1538 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF1538
AF1538-SP

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Mouse Cripto Antibody Summary

Species Reactivity

Mouse

Specificity

Detects mouse Cripto in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross-reactivity with recombinant human Cripto-1 is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

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 either lyophilized or 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

Western Blot

0.1 µg/mL

Recombinant Mouse Cripto (Catalog # 1538-CR)

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 μg/mL, this antibody will block >90% of the binding.

Immunocytochemistry

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.

Scientific Data

Immunocytochemistry

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Cripto in D3 Mouse Cell Line. Cripto was detected in immersion fixed D3 mouse embryonic stem cell line using Sheep Anti-Mouse Cripto Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1538) 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 and cell secretion. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Reconstitution Calculator

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: 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

Rosa, F.M. (2002) Science’s STKE http://stke.sciencemag.org/.
Shen, M. and A. Schier (2000) Trends Genet. 16:303.
Yan, Y-T. et al. (2002) Mol. Cell Biol. 22:4439.
Salomon, D. et al. (2000) Endocrine-Rel. Cancer 7:199.
Gray, P.C. et al. (2003) Proc. Natl. Acad. Sci. USA 100:5193.
Cheng, S. et al. (2003) Genes & Dev. 17:31.
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