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Human Cripto-1 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
62.5 - 4,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Cripto-1. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008B) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Block Buffer: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Reagent Diluent: 0.1% BSA, 0.05% Tween 20 in Tris-buffered Saline (20 mM Trizma base, 150 mM NaCI) pH 7.2-7.4, 0.2 μm filtered

Substrate Solution: TMB ELISA Substrate (Catalog # DY999B)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

 

*For the Reagent Diluent and Blocking Buffer recommended for a specific DuoSet ELISA Development Kit, please see the product.

Data Example

Human Cripto 1 ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Cripto

Cripto, also known as TDGF-1 (Teratocarcinoma-Derived Growth Factor), is the founding member of the Epidermal Growth Factor-Cripto-1/FRL1/Cryptic Protein family of signaling proteins that function in cancer and various developmental processes. Mouse Cripto shares 66% and 34% amino acid identity with human Cripto and zebrafish one-eyed pinhead (OEP), respectively. Genetic evidence from mice and zebrafish points to a role for Cripto as an essential cofactor in Nodal signaling.

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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human Cripto-1 DuoSet ELISA

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.

4 Citations: Showing 1 - 4
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  1. Oncofetal Protein CRIPTO Is Involved in Wound Healing and Fibrogenesis in the Regenerating Liver and Is Associated with the Initial Stages of Cardiac Fibrosis
    Authors: S Karkampoun, D van der He, M Scarpa, B van Hoek, HW Verspaget, MJ Goumans, MJ Coenraad, BPT Kruithof, M Kruithof-d
    Cells, 2021;10(12):.
    Species: Human
    Sample Types: Plasma
  2. CRIPTO and miR-371a-3p Are Serum Biomarkers of Testicular Germ Cell Tumors and Are Detected in Seminal Plasma from Azoospermic Males
    Authors: CM Spiller, J Lobo, WPA Boellaard, AJM Gillis, J Bowles, LHJ Looijenga
    Cancers (Basel), 2020;12(3):.
    Species: Human
    Sample Types: Serum
  3. Expression of Cripto-1 predicts poor prognosis in stage I non-small cell lung cancer
    Authors: C Xu, Q Yuan, H Hu, W Wang, Q Zhang, L Li, J Wang, R Yang
    J. Cell. Mol. Med., 2020;0(0):.
    Species: Human
    Sample Types: Serum
  4. 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;22(10):1700-13.
    Species: Human
    Sample Types: Cell Culture Supernates

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