Custom human Cripto-1 ELISA Kit

Product Features

Product Name           Custom human Cripto-1 ELISA Kit
Assay Type                   Solid Phase Sandwich ELISA
Format                           96-well strip plate
Assay Length              4.5 hours
Sample Volume         Serum ,Plasma, cell culture media , 100ul
Detection Method    Colorimetric
Sensitivity                   This has not yet been determined
Assay Range                This has not yet been determined
Reactivity                     Human
Cross-Reactivity       This ELISA antibody pair only recognizes human Cripto-1
Gene Names:              TDGF1, CR, CRGF, CRIPTO
Gene ID                         6997
Swissprot ID               P13385

Kit Components

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent
• Standards
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Protocol Outline

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 µl of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4°C.
4. Wash and add 100 µl of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Wash and add 100 µl of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Wash and add 100 µl of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 µl of Stop Solution to each well.
11. Read at 450 nm/570 nm.

Background

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. 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. 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. Both secreted and membrane bound forms of Cripto demonstrate biological activity. Cripto and OEP mutants display defects in mesoderm induction and heart morphogenesis, similar to phenotypes seen in Nodal mutants. Cripto also forms a complex with activin and Act RIIs to block activin signaling. 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.

Storage

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Regulatory Status      

Custom human Cripto-1 ELISA Kit is for Research Use Only.

• Manual
• Assay procedure
• MSDS