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, 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.
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.
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).
10 – 100µg/ml
10µg/ml
Colorimetric Detection (655nm) (Endpoint)
100 in total (allows a maximum of 46 samples to be run in duplicate alongside a standard curve).
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.
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.
Mode of ActionAssay SpecificationsKit 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.
Description
The ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) is a ready-to-use reagent with all the essential components for quantitative real-time PCR (qPCR) except primers and templates. The master mix contains a highly stable hot-start Taq polymerase in an optimized buffer with dsDNA specific SYBR green fluorescent dye. Consequently, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) features high stability during storage, even at 37°C for weeks. In addition to high sensitivity and signal intensity, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) performs a low background/ high specificity qPCR results, as well as a better compatibility with fast PCR program.
With inert smart blue contrast dye, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) is ready-to-use and greatly reduces pipetting errors, while largely improving the reproducibility of the process. The ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) is also compatible with ROX reference dye if ROX is recommended by the manufacturer of the qPCR system. This master mix allows sensitive, precise amplification, real-time tracking of the amplification process, and simultaneous quantification for targeted DNA molecules.
Features
Storage
Protect from light.
Aliquot to avoid multiple freeze-thaw cycles.
-20°C for 12 months
The ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) is a ready-to-use reagent with all the essential components for quantitative real-time PCR (qPCR) except primers and templates. The master mix contains a highly stable hot-start Taq polymerase in an optimized buffer with dsDNA specific SYBR green fluorescent dye. Consequently, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) features high stability during storage, even at 37°C for weeks. In addition to high sensitivity and signal intensity, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) performs a low background/ high specificity qPCR results, as well as a better compatibility with fast PCR program.
With inert smart blue contrast dye, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) is ready-to-use and greatly reduces pipetting errors, while largely improving the reproducibility of the process. The ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, no ROX) is also compatible with ROX reference dye if ROX is recommended by the manufacturer of the qPCR system. This master mix allows sensitive, precise amplification, real-time tracking of the amplification process, and simultaneous quantification for targeted DNA molecules.
The Hepatitis B virus (HBV) is mainly transmitted via blood or blood products. In addition, sexual, oral and perinatal infections are also possible. Early symptoms of the infection include appetite loss, vomiting and abdominal symptoms, with approximately 10-20% of those patients developing fever as well as rheumatoid joint and muscle pain. Jaundice, which may be accompanied by itching, will then develop within 2-14 days. Fulminant hepatitis then occurs in about 1% of all infected patients, which in severe cases may be fatal. Of those individuals infected by HBV, 5-10% will develop chronic liver inflammation which may progress to cirrhosis of the liver or, in the worst case, primary liver cell carcinoma. The HBV virus is a hepadnavirus that has a circular genome composed of partially double-stranded DNA.
HBV TaqMan PCR Kit, 100 reactions
HBV TaqMan PCR Probe/Primer Set and Controls, 100 reactions
For research use only and NOT intended for in vitro diagnostics.
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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival.
| Component | Cat. TM29250 (100 rxns) | Cat. TM29210 (100 rxns) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| HBV Primer & Probe Mix | 280 μL | 280 μL |
| HBV Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |