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.

Description

•  Appearance: White/pink
• • Length of strip: 86mm
• • Width of strip:  4.5mm
• • Length of device: 136mm
• • Width of device: 16mm

Introduction

A highly efficient, easily automated agarose gel or PCR purification system that delivers superior quality DNA with no salt carryover. Requiring no centrifugation or filtration. The Kit can be easily used in manual and automated 96 or 384-well formats.

Details

Specifications

Features	Specifications
Main Functions	Recover DNA fragments >100bp from agarose gel(<0.3g), purification of DNA from PCR, enzymatic reaction solution or crude gDNA
Applications	PCR, NGS, chip analysis, etc.
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Agarose Gel, PCR products, enzymatic reaction solution
Sample amount	Agarose Gel: <0.3g,  PCR products: ≤50µl
Recovery	80%
Elution volume	≥30μl
Time per run	≤30 minutes

The Kit method contains magnetic particles in an optimized binding buffer to selectively bind DNA fragments (>100bp) and larger to paramagnetic beads. Excess primers, nucleotides, salts, and enzymes can be removed using a simple washing procedure.

Advantages

• High recovery efficiency – up to 80% DNA recovery
• General – recover DNA from gel or enzyme-driven reaction solutions such as PCR

Kit Contents

Contents	D500101	D500102	D500103
Purification Times	50 Preps	500 Preps	5000 Preps
Buffer GDP	30 ml	250 ml	3 x 900 ml
MagPure Particles	1.6 ml	15 ml	3 x 50 ml
Buffer DW1	20 ml	180 ml	2 x 800 ml
Elution Buffer(10mmTris, pH8.5)	10 ml	60 ml	500 ml

Storage and Stability

MagPure Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

Experiment Data