Human Cryptic Antibody

Catalog # Availability Size / Price Qty
AF1410
AF1410-SP
Detection of Cryptic in iPSC mesoderm differentiated cells by Flow Cytometry.
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Human Cryptic Antibody Summary

Species Reactivity
Human
Specificity
Detects human Cryptic in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant mouse Cryptic is observed.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Cryptic
Tyr26-Gly169
Accession # NP_115934
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human Cryptic (Catalog # 1410-CR)
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Intracellular Staining by Flow Cytometry
0.25 µg/106 cells
iPSC mesoderm differentiated cells treated with iPSC kit for Mesoderm differentiation (Catalog # SC030B)

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

Intracellular Staining by Flow Cytometry View Larger

Detection of Cryptic in iPSC mesoderm differentiated cells by Flow Cytometry. iPSC mesoderm differentiated cells treated with iPSC kit for Mesoderm differentiation (SC030B) were stained with Sheep Anti-Human Cryptic Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1410, filled histogram) or isotype control antibody (Catalog # 5-001-A, open histogram), followed by Phycoerythrin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0126). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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: Cryptic

Cryptic, also known at CFC-1, was originally identified as a secreted protein that is specifically expressed during mesoderm differentiation (1). Cryptic, along with Cripto, Xenopus FRL-1 and zebrafish OEP (one-eyed pinhead) define the epidermal growth factor-CriptoFRL1Cryptic (EGF-CFC) family of signaling proteins that function in cancer and various developmental processes (2, 3). Overall sequence identity between members of the family is low, but they do share distinct domains: a variant EGF-like motif, a novel conserved cysteine-rich domain (called CFC domain), and a C-terminal hydrophobic region (2). 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 Cryptic shares 55% and 25% amino acid idenitity with mouse Cryptic and human Cripto, respectively. 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).

Cryptic is expressed during gastrulation in the mesoderm and later in the neuroectoderm, marking the prospective floor plate of the neural tube (1). Genetic evidence from mice and humans points to a role for Cryptic in determining left-right asymmetry. Mutations in the cryptic gene result in a spectrum of heart, lung and spleen defects, all representing left-right laterality defects (4, 5). These phenotypes resemble some Nodal mutant alleles suggesting that Cryptic, like Cripto, acts as an essential co-factor for Nodal signaling (1, 3). Studies have shown that other TGF-beta superfamily members involved in mesoderm induction and left-right patterning, Vg1 and GDF-1, also require EGF-CFC co-factors. Cryptic binds to GDF-1 leading to an Act RIIB-ALK4-Cryptic-GDF-1 complex for signaling (6).

References
  1. Shen, M. et al. (1997) Development 124:429. 
  2. Shen, M. and A. Schier (2000) Trends Genet. 16:303.
  3. Rosa, F.M. (2002) Science's STKE http://stke.sciencemag.org/.
  4. Gaio, U. et al. (1999) Curr. Biol. 9:1339.
  5. Bamford, R. et al. (2000) Nature Genet. 26:365.
  6. Cheng, S. et al. (2003) Genes & Dev. 17:31.
Entrez Gene IDs
55997 (Human); 12627 (Mouse)
Alternate Names
CFC1; CFC-1; CFC1B; cripto, FRL-1, cryptic family 1; Cryptic family protein 1; cryptic protein; Cryptic; CRYPTICFLJ77897; HTX2MGC133213

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