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A competitive ELISA-like kit using an HA binding protein optimized to quantify HA polymers of average molecular weight greater than 7.4 kDa in samples such as serum, plasma and culture supernatant.

Hyaluronan Quantification Kit

Background

Hyaluronan (HA) is an unbranched glycosaminoglycan composed of repeating disaccharide units of D-glucronic acid and N-acetyl-D-glucosamine. HA is abundant in synovial fl uid, skin, umbilical cord, and vitreous bodies. HA is a prominent component of the extracellular matrix and contributes to tissue water retention and to cellular growth, diff erentiation, and migration. HA exists in the body in a wide range of molecular weights (MW) derived from several distinct HA synthases and degradation enzymes. For example, the average molecular weights of HA in synovial fl uid, blood and urine are 4-6 MDa, 100-300 kDa and less 10 kDa, respectively.

This product is a competitive ELISA-like kit using an HA binding protein optimized to quantify HA polymers of average molecular weight greater than 7.4 kDa in samples such as serum, plasma and culture supernatant.

Measurement principle Fig. 1 Measurement principle
A sample containing HA and Biotin-HABP are added to an HA-coated plate. HRP-Avidin is then added, and the HA binding Biotin-HABP is detected by chromophoric substrate (Competition Method).

Features

* Hardly getting aff ected by the molecular size of HA
* Measurable the HA concentrations accurately in blood serum, plazma and supernatant of which the molecular weight is above 7.4 kDa.
* Measuring range of HA concentration is between 3.13 - 200 ng/mL.

Kit Components

Description Volume Quantity
96 well plate 8 x 12 wells 1
HA Coating Solution 11 mL 1
Wash Buffer (20X) 50 mL 1
Sample Buffer (2X) * 50 mL 1
Blocking Buffer (2X) * 11 mL 1
HA Standard (10 µg/mL) * 0.1 mL 1
96 well plate 8 x 12 wells 1
Biotin-HABP * 10 - 40 µL 1
HRP-Avidin (100X) * 0.2 mL 1
Substrate Solution A 1 mL 1
Substrate Solution B 10 mL 1
Stop Solution 11 mL 1
Plate Seals 5 mL 2
96 well plate 8 x 12 wells 1

* : containing 0.025 - 0.05% Proclin 300

Measuring Range

Typical calibration curve
The measuring range of HA using Hyaluronan Quantification Kit is 3.13~200 ng/mL.

Product Data: Specificity

The reactivity with various molecular weights of HA (HA with molecular weights of from 2.8 to 2367 kDa were compared)
The reactivity with various molecular weights of HA was evaluated using Hyaluronan Quantification Kit . When using HA with molecular weights of from 2.8 to 2367 kDa, this kit reacted with molecular weights of over 7.4 kDa HA in a similar manner. HA with a molecular weight of 2.8 kDa showed a slightly weaker reactivity.

The cross-reactivity with various kind
The cross-reactivity with various kinds of glycosaminoglycans (GAGs) was evaluated using Hyaluronan Quantification Kit. This kit did not show the cross-reactivity with chondroitin sulfate-A (CS-A), CS-C, CS-D, CS-E, heparan sulfate (HS), heparin (Hep), keratan sulfate (KS) and keratan polysulfate (KPS) at the concentrations of up to 1000 ng/mL. A weak reactivity was shown in high concentration of CS-B (dermatan sulfate).

HA contents in plasma and serum

Reagent Preparation and Assay Procedure

Please refer Manual and Technical data

Product List

Product Name Cat# Quantity Price

Hyaluronan Quantification Kit

CSR-HA-96KIT 1KIT

¥ 70,000
$ 934
€ 700

References
  • Haserodt S et al., A comparison of the sensitivity, specifi city, and molecular weight accuracy of three diff erent commercially available Hyaluronan ELISA-like assays. Glycobiology. 21(2):175-183. 2011.
    PMID: 20864567
  • Maeda H et al., A competitive enzyme-linked immunosorbent assay-like method for the measurement of urinary hyaluronan. Biosci Biotechnol Biochem, 63(5):892-895, 1999.
    PMID: 10380631

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.