Purple-Jelley Hyaluronic Acid assay kit

Biocolor’s Purple-Jelley assay kit is the perfect tool for accurate measurement of hyaluronic acid / Hyaluronan levels in your samples. This colorimetric assay is optimised for quantitative analysis in-vivo, tissue-derived hyaluronic acid / Hyaluronan and includes full step-by-step instructions.

Colorimetric Detection (655nm) (Endpoint)

Hyaluronic Acid: A gentle giant!

Hyaluronic acid, in its hydrated form, is a unique carbohydrate polymer, often referred to as a ‘gentle giant.’ It consists of a lengthy, flexible, non-branching chain with a repeating disaccharide pattern. This disaccharide is composed of alternating uronic acid and aminosugar units.

Why is our kit called ‘Purple-Jelley’?

Discovering the J-Aggregate Effect in Cyanine DyesIn 1936, Edwin Jelley made a fascinating observation, documented it in a letter to Nature (Nature 138, 1009 – 1010). He noted a peculiar behaviour of certain cyanine dyes, that when dissolved in 5 M NaCl, they dyes exhibited a third absorbance peak at a longer wavelength, around 650nm. In deionized water, however, they displayed only a double peak at approximately 540nm and 570nm. The 650nm peak in concentrated dye solutions resulted from the aggregation of dye molecules and was later termed a ‘J-aggregate,’ in honor of Edwin Jelley. The J-aggregate is known as a supra-molecular complex, formed by stacking individual dye molecules.

Subsequent research in the 1960s, notably by Kay et al. (J. Physical Chem. 68, 1896 – 1906), revealed that various biological polymers, including proteins, DNA, polar lipids, and glycosaminoglycans, could also induce this third absorbance peak. This phenomenon led to the development of the Purple-Jelley assay, named after the purple color of the dye reagent and Edwin Jelley himself.

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An overview of the Purple-Jelley assay steps:

During the assay, hyaluronic acid is selectively purified during the assay sample preparation protocol. This is then reacted with the Purple-Jelley dye reagent, and the absorption of the characteristic third wavelength recorded. By comparison with a calibration curve the hyaluronic acid content of the sample can be measured.

‍Step 1. The assay protocol takes tissue samples through a sequential sample preparation protocol which involves enzymatic protein digestion, followed by precipitation and purification of GAGs, culminating in the precipitation of purified Hyaluronic acid.

Step2. The processed sample is then incubated for 10 minutes with the Purple-Jelley dye reagent, forming a coloured product which can be measured spectrophotometrically.

Step 3. The Hyaluronic acid content of unknown samples can be calculated by comparison against a calibration curve prepared using a standard comprising hyaluronic acid (supplied with the kit).

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Assay range

10 – 100µg/ml

Limit of Detection

10µg/ml

Detection Method

Colorimetric Detection (655nm) (Endpoint)

Measurements per kit

100 in total (allows a maximum of 46 samples to be run in duplicate alongside a standard curve).

Suitable Samples

In-vivo: Hyaluronic acid purified from in-vivo tissues. The kit protocol involves extraction and purification of hyaluronic acid prior to reaction with the Purple-Dye reagent.

Precautions

This kit is designed for research use only. Not for use in diagnostic procedures.
Kit requires access to a centrifuge, as well as a spectrophotometer/colorimeter capable of colorimetric, absorbance detection at 655nm.
Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.

ation

Mode of ActionAssay SpecificationsKit Contents

Purple-Jelley Hyaluronic acid kit contents:‍

1. Purple-Jelley Dye Reagent (1x 20ml)

2. Hyaluronan Reference Standard (1x 5ml, 0.2mg/ml soluble Hyaluronic Acid)

3. Precipitating Reagent (2x 34ml)

4. Sodium Chloride (1x 20ml)

5. Cetylpyridinium Chloride (1x 20ml)

6. TRIS-buffered Saline (5x tablets)

7. 2ml screw-cap tubes for preparation of samples.

8. Assay kit manual

NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details.

Biocolor’s Purple-Jelley assay kit is the perfect tool for accurate measurement of hyaluronic acid / Hyaluronan levels in your samples. This colorimetric assay is optimised for quantitative analysis in-vivo, tissue-derived hyaluronic acid / Hyaluronan and includes full step-by-step instructions.

Overview

• Reliable and cost-effective
• Non-invasive, user-friendly sample collection and preservation in one convenient kit
• Preserved RNA is stable for up to 2 months at room temperature
• Compatible with most RNA isolation methods including Norgen’s Saliva/Swab RNA Purification Kits
• High quality RNA is suitable for sensitive downstream applications including RT-PCR, RT-qPCR, RNA sequencing and microarray
• Inactivates viruses including SARS-CoV-2 upon mixing – Explore the Application Note
• Shipping accessories can be purchased separately
• CE-IVD marked version available

Norgen’s Saliva RNA Collection and Preservation Devices are designed for simple and non-invasive saliva collection and preservation of RNA in saliva samples at ambient temperature. Each of the 50 Saliva RNA Collection and Preservation Devices consists of 3 components:

• Saliva Collection Funnel and Collection Tube
• Collection Tube Cap
• Norgen’s Saliva RNA Preservative contained within a sealed squeezable ampoule

Saliva samples are collected by spitting inside the Collection Funnel which has been assembled with the Collection Tube. After collecting the required volume of saliva the Collection Funnel is removed and the contents of the Preservative Ampoule are then added and mixed with the collected saliva. The Saliva Collection Tube is subsequently sent to the laboratory for RNA isolation and analysis. RNA can be isolated from the preserved saliva samples using Norgen’s Saliva/Swab RNA Purification Kits (Cat# 69100, 69300). Each of Norgen’s Collection Tubes is labeled with a unique serial number that can be used for secure and anonymous tracking of the sample. The Saliva RNA in preserved samples is stable for up to 2 months at room temperature. This kit is ideal for collecting and preserving RNA samples for epidemiological and population studies.

Saliva RNA Preservative

Norgen’s Saliva RNA Preservative is an aqueous storage buffer designed for rapid cellular lysis and subsequent preservation of RNA from fresh specimens. The buffer eliminates the need to immediately process or freeze samples and allows the samples to be shipped to centralized testing facilities at ambient temperature. The components of the buffer allow samples to be stored for up to 2 months at room temperature.

Nucleic Acid Isolation from Preservative

Saliva RNA can be isolated from the preserved saliva samples using Norgen’s Saliva/Swab RNA Purification Kits (Cat# 69100, 69300).

Details

Supporting Data

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Kit Specifications
Volume of Saliva Collected	2 mL
Volume of Saliva-Preservative Mix	4 mL
Preservation Temperature	Room Temperature
Preservation time	Up to 2 months at room temperature

Shelf Life and Handling

• When stored at room temperature, unused Norgen Saliva Preservative Tubes are stable through to the collect-before date without any reduction in performance. Please see the device insert or tube label for collect-before date.
• Once collected, saliva RNA is stable for up to 2 months when kept tightly sealed and stored at room temperature.
• The Collection Tube, the Collection Funnel and the Device Container of each individual Saliva RNA Collection and Preservation Device are recyclable.

Kit Components	Cat. RU53800 (50 Devices)
Individual Saliva RNA Collection and Preservation Devices	50
Donor Procedure Flowchart	1
Product Insert	1
Individual Saliva RNA Collection and Preservation Device Contents
Saliva Collection Funnel and Collection Tube	1
Collection Tube Cap	1
Preservative Ampoule	1
Donor Instructions	1