Hyaluronan, also known as hyaluronic acid (HA) or sodium hyaluronate, is a naturally occurring linear polymer. Hyaluronan is a glycosaminoglycan (GAG) that is ubiquitously present in the extracellular matrix of all vertebrates and is also present in the capsule of some strains of Streptococci. Mammalian hyaluronan is synthesized by one of three distinct hyaluronan synthases (HAS1, 2, and 3), which produce HA polymers with different chain lengths and differ in their rates of synthesis. Whereas high molecular weight HA (> 500 kDa) is anti-angiogenic, anti-inflammatory and immunosuppressive, low molecular weight HA (10 - 500 kDa) is highly angiogenic and pro-inflammatory. HA oligomers are anti-apoptotic and upregulate heat shock protein expression. Functionally, hyaluronan molecules are important for the maintenance of a highly hydrated extracellular matrix in tissues, which is involved in cell adhesion and supports cell migration.
Hyaluronan DuoSet ELISA
R&D Systems | Catalog # DY3614
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
Hyaluronan DuoSet ELISA Features
- Optimized capture and detection reagent 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
- Available in 5 (96-well) plate pack size
- Economical alternative to complete kits
Product Summary for Hyaluronan DuoSet ELISA
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural Hyaluronan. 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 Specifications
Assay Format
Sample Volume Required
Detection Method
Conjugate
Specificity
Label
Scientific Data Images for Hyaluronan DuoSet ELISA
Hyaluronan ELISA Standard Curve
Kit Contents for Hyaluronan DuoSet ELISA
- Capture Reagent
- Detection Reagent
- Natural Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 3 (5 plates): (Catalog # DY009C) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 3.
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: Hyaluronan
Additional Hyaluronan Products
Product Documents for Hyaluronan 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 Hyaluronan DuoSet ELISA
For research use only
Related Research Areas
Citations for Hyaluronan DuoSet ELISA
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Customer Images
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Sample Tested: Supernatant Human Epidermal keratinocytes (HEKa) in presence or absence of EGF and Cultured Human KeratinocytesVerified Customer | Posted 06/22/2023Hyaluronan DuoSet ELISA standard curve
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Sample Tested: Cell culture supernatantVerified Customer | Posted 11/03/2021
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Sample Tested: Cell culture supernatantVerified Customer | Posted 12/15/2020
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Protocols
View specific protocols for Hyaluronan DuoSet ELISA (DY3614):
GENERAL ELISA PROTOCOL
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 of 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.
Assay Procedure
- 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.
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for Hyaluronan DuoSet ELISA
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Q: Can you tell me if the Hyaluronan DouSet ELISA will is suitable to use with pig plasma?
A: Yes, this ELISA is for any species.