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.

Introduction

This product supplies a simple and rapid extraction of total RNA from tissue and culture cells samples. The kit is based on superparamagnetic particles purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction processis only 60 minutes. Purified RNA is ready for downstream applications such as RT-PCR, virus RNA testing and so on. MagPure LQ Kit buffers can be used for both manual extraction process and automatic nucleic acid extraction machines.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from tissue and cells and plant
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor, pipetting workstation
Sample type	Animal tissue, human tissue, cultured cells, lymphocytes, ordinary plant tissue
Sample amount	Cells: 1 x 107Animal tissue: ≤20mgPlant tissue: ≤100mgYeast cell: ≤5 x 106
Yield	5-100μg

Principle

The Kit combines the speed and efficiency of silica-based technology with the convenient handling of magnetic particles for purification of total RNA. Samples are lysed and RNA is purified from lysates in one step through its binding to the silica surface of the particles in the presence of a chaotropic salt. The particles are separated from the lysates using a magnet and DNA is removed by treatment with RNase-free DNase. The magnetic particles are efficiently washed, and RNA is eluted in RNase-free water.

Advantages

• High purity – OD 260 / 280 :1.9-2.0, OD 260 / 230 :1.5-2.0
• Economy – less than 50% of the price of Qiagen and other imported products
• Automatic – extraction without manual participation, saving time and effort
• Universal – suitable for various animal tissues, cultured cells and conventional plant fungal tissues

Kit Contents

Contents	R662201	R662202	R662203
Purification Times	48 Preps	96 Preps	5 x 96 Preps
MagPure RNA Particles	1.7 ml	4.0 ml	18 ml
Proteinase K	24 mg	50 mg	240 mg
Protease Dissolve Buffer	1.8 ml	5 ml	15 ml
DNase I	600 μl	2 x 600 μl	10 x 600 μl
DNase Buffer	30 ml	40 ml	200 ml
RTL Lysis Buffer	40 ml	80 ml	400 ml
Buffer MCB*	18 ml	30 ml	150 ml
Buffer MW1*	44 ml	66 ml	2 x 220 ml
Buffer RW2*	20 ml	50 ml	2 x 100 ml
RNase Free Water	10 ml	30 ml	120 ml

Storage and Stability

MagPure RNA Particles and Proteinase K should be stored at 2–8°C upon arrival. DNase I shouldbe stored at -20°C. However, short-term storage (DNase I up to 1 weeks, MagPure RNA Particles and Proteinase K up to 8 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 atleast 18 months under these conditions.

This product supplies a simple and rapid extraction of total RNA from tissue and culture cells samples. The kit is based on superparamagnetic particles purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction processis only 60 minutes. Purified RNA is ready for downstream applications such as RT-PCR, virus RNA testing and so on. MagPure LQ Kit buffers can be used for both manual extraction process and automatic nucleic acid extraction machines.

Introduction

MagPure Stool DNA Kit is specially designed for high throughput DNA extraction from stool samples. It can get high purity microbial DNA from stool samples (≤200mg). This kit is based on magnetic beads purification and unique inhibiting factors adsorption technology, no phenol-chloroform extraction or alcohol precipitation. It can adsorb humic acid and other inhibiting factors in the solution efficiently. DNA can be directly used for downstream applications such as PCR, Viral DNA testing, bacterial DNA testing, etc.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 100-150mg stool samples
Applications	PCR, southern blot and enzyme digestion, etc.
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Stool
Sample amount	100-150 mg
Elution volume
Time per run

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• Fast – several samples can be extracted in 60 minutes (after digestion)
• High purity – unique adsorbent can completely remove inhibitory factors
• High recovery – DNA can be recovered at the level of PG

Kit Contents

Contents	D636401	D636402	D636404
Purification Times	48 Preps	96 Preps	400 Preps
MagPure Particles N	1.7 ml	3.4 ml	14 ml
2ml Bead Tubes	48	96	400
RNase A	10 mg	20 mg	75 mg
Proteinase K	24 mg	48 mg	180 mg
Protease Dissolve Buffer	3 ml	5 ml	20 ml
Buffer ATL	60 ml	110 ml	300 ml
PVP-10	1.2 g	2.2 g	6 g
Buffer PCI	50 ml	100 ml	300 ml
Buffer MLE	30 ml	60 ml	180 ml
Buffer GW1*	22 ml	44 ml	132 ml
Elution Buffer	20 ml	20 ml	60 ml

Storage and Stability

MagPure Particles, RNase A and Proteinase K 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 dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

MagPure Stool DNA Kit is specially designed for high throughput DNA extraction from stool samples. It can get high purity microbial DNA from stool samples (≤200mg). This kit is based on magnetic beads purification and unique inhibiting factors adsorption technology, no phenol-chloroform extraction or alcohol precipitation. It can adsorb humic acid and other inhibiting factors in the solution efficiently. DNA can be directly used for downstream applications such as PCR, Viral DNA testing, bacterial DNA testing, etc.