Human/Primate Cripto-1 Antibody

Catalog #: MAB2772
1 Citation Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB2772-SP
MAB2772-100
MAB2772-500

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Primate Cripto-1 Antibody Summary

Species Reactivity

Human, Primate

Specificity

Detects human and primate Cripto-1 in ELISAs. In sandwich immunoassays, no cross-reactivity with recombinant mouse Cripto-1, recombinant human (rh) EGF, rhTGF‑ alpha, rhTGF-beta 1, rhTGF-beta 2, rhTGF-beta 3, or rhCryptic is observed.

Source

Monoclonal Mouse IgG₁ Clone # 89633

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human Cripto-1
Arg38-Tyr188
Accession # P13385

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

Flow Cytometry

0.25 µg/10⁶ cells

BG01V human embryonic stem cells

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.1-0.5 µg/mL of this antibody will block 50% of the binding of 75 ng/mL of Recombinant Human Cripto-1 (Catalog # 145-CR) to immobilized Recombinant Human Activin RIB/ALK-4 Fc Chimera (Catalog # 808-AR) coated at 2 µg/mL (100 µL/well). At 20 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

5-25 µg/mL

See below

Human/Primate Cripto-1 Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

2-8 µg/mL

Use in combination with:

Detection Reagent: Human/Primate Cripto-1 Biotinylated Antibody (Catalog # BAM2773)

Standard: Recombinant Human Cripto-1 Protein (Catalog # 145-CR)

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-1 in Human iPSCs. Cripto-1 was detected in immersion fixed human induced pluripotent stem cells (iPSCs) using Mouse Anti-Human/Primate Cripto-1 Monoclonal Antibody (Catalog # MAB2772) at 20 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Stem Cells on Coverslips.

Flow Cytometry

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Detection of Cripto-1 in BG01V Human Cells by Flow Cytometry. BG01V human embryonic stem cells were stained with Mouse Anti-Human Cripto-1 Monoclonal Antibody (Catalog # MAB2772, filled histogram) or isotype control antibody (MAB002, open histogram) followed by anti-Mouse IgG APC-conjugated Secondary Antibody (F0101B). Staining was performed using our Staining Membrane-associated Proteins protocol.

Reconstitution Calculator

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. Human Cripto shares 66% and 28% amino acid identity with mouse 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). Studies have 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

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Citation for Human/Primate Cripto-1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

Dynamic regulation of the cancer stem cell compartment by Cripto-1 in colorectal cancer.
Authors: Francescangeli F, Contavalli P, De Angelis M, Baiocchi M, Gambara G, Pagliuca A, Fiorenzano A, Prezioso C, Boe A, Todaro M, Stassi G, Castro N, Watanabe K, Salomon D, De Maria R, Minchiotti G, Zeuner A
Cell Death Differ, 2015-03-20;22(10):1700-13.
Species: Human
Sample Types: Cell Culture Supernates
Applications: ELISA Development (Capture)