Human CEACAM-5/CD66e DuoSet ELISA

R&D Systems | Catalog # DY4128

R&D Systems
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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

31.2-2000 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Human

Human CEACAM-5/CD66e DuoSet ELISA 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
View all CEACAM-5/CD66e ELISA Kits »

Product Summary for Human CEACAM-5/CD66e DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human CEACAM-5 / CD66e. 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 ELISA.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

100 µL

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human CEACAM-5/CD66e DuoSet ELISA

Human CEACAM-5 / CD66e ELISA Standard Curve

Human CEACAM-5 / CD66e ELISA Standard Curve

Kit Contents for Human CEACAM-5/CD66e DuoSet ELISA

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

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) 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

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

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

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: CEACAM-5/CD66e

CEACAM-5, also known as CEA and CD66e, belongs to the large family of CEACAM and pregnancy specific glycoproteins. CEACAM molecules are either transmembrane or GPI-linked, and are differentially expressed between species. Orthologs of human CEACAM-5 have not been described in other species. CEACAM-5, which is expressed primarily by epithelial cells, consists of an N-terminal Ig like V-set domain followed by six Ig-like C2-set domains and a GPI anchor. CEACAM-5 is synthesized as a 180 kDa, variably glycosylated molecule of which approximately 60% is carbohydrate. CEACAM-5 functions as a calcium independent adhesion molecule through homophilic and heterophilic interactions with CEACAM-1.

CEACAM-5 is restricted to the apical face of intestinal epithelial cells in the adult but is more diffuse during embryonic development and in tumors. This is consistent with a role in the development and maintenance of epithelial architecture. CEACAM-5 is upregulated in a wide variety of human tumors and is a commonly used cancer marker. It promotes tumor cell migration, invasion, adhesion, and metastasis. CEACAM-5 also contributes to tumor formation by maintaining cellular proliferation in the presence of differentiation stimuli, and by blocking apoptosis following loss of ECM anchorage (anoikis).

Long Name

Carcinoembryonic Antigen-related Cell Adhesion Molecule 5

Alternate Names

CD66e, CEA, CEACAM5

Entrez Gene IDs

1048 (Human); 73250 (Mouse); 102128791 (Cynomolgus Monkey)

Gene Symbol

CEACAM5

Additional CEACAM-5/CD66e Products

Product Documents for Human CEACAM-5/CD66e DuoSet ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CEACAM-5/CD66e DuoSet ELISA

For research use only

Citations for Human CEACAM-5/CD66e DuoSet ELISA

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    Human CEACAM-5/CD66e DuoSet ELISA DY4128

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Protocols

View specific protocols for Human CEACAM-5/CD66e DuoSet ELISA (DY4128):

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.

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