CD96 (also Tactile) is a 160 kDa member of the Ig-Superfamily. It is expressed on CD4+ and CD8+ T cells, plus NK cells and select B cells. Human CD96 binds to CD155 and presumably participates in NK cell killing of CD155-expressing target cells. Mature human CD96 is a 564 amino acid (aa), type I transmembrane glycoprotein. It contains a 498 aa extracellular region (aa 22-519) that contains three Ig-like domains. The two N-terminal domains are V-type (aa 38-238), while the distal domain is a C-type structure (aa 269-375). There is one isoform that shows a deletion of aa 183-192. This deletion converts the second V-type domain into an I-like domain, and generates the most common form of CD96. An additional isoform shows the same deletion coupled to a nine aa substitution for aa 410-585. Over aa 1-536, human CD96 shares 59% aa identity with mouse CD96.
Key Product Details
Assay Type
Assay Range
Sample Type
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Reactivity
Human CD96v2 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
Product Summary for Human CD96v2 DuoSet ELISA
Product Specifications
Assay Format
Detection Method
Conjugate
Specificity
Label
Scientific Data Images for Human CD96v2 DuoSet ELISA
Human CD96v2 DuoSet Standard Curve
Kit Contents for Human CD96v2 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 # DY008C) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
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 equivalent
Reagent Diluent*
Blocking Buffer*
Substrate Solution: ELISA TMB Substrate (Catalog # DY999B or DY999B-250)
Stop Solution: Methanesulfonic acid (Catalog # DY994B or DY994B-250)
Microplates: (Catalog # DY990), or equivalent
Plate Sealers: (Catalog # DY992), or equivalent
*For the recommended Reagent Diluent and Blocking Buffer for a specific DuoSet ELISA Development Kit, refer to the product datasheet.
Preparation and Storage
Shipping
Stability & Storage
Background: CD96
Alternate Names
Gene Symbol
Additional CD96 Products
Product Documents for Human CD96v2 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.
Product Specific Notices for Human CD96v2 DuoSet ELISA
For research use only
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Protocols
View specific protocols for Human CD96v2 DuoSet ELISA (DY61991-05):
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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.
Associated Pathways
