Introduction

This product is suitable for rapid RNA extraction from tissue , cells, and other clinical samples. RNA can be used directly for RT-PCR, quantitative RT-PCR and so on.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from tissue, cell
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	Tissues, cells, lymphocytes and other clinical sample
Sample amount	Cells grown in suspension:3~5 x 106Animal tissue: 10~20mgPlant tissue: ≤100mg

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 I. The magnetic particles are efficiently washed, and RNA is eluted in RNase-free water

Advantages

• High purity – OD260 / 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 clinical samples

Kit Contents

Contents	IVD3020
Purification Times	200 Preps
MagPure RNA Particles	7 ml
DNase I	4 x 600 µl
DNase Buffer	80 ml
RTL Lysis Buffer	150 ml
Buffer MCB*	75 ml
Buffer MW1*	110 ml
Buffer MW2*	50 ml
RNase Free Water	60 ml

Cat.No	Reagent	IVD3020-F-96
DNase Buffer		60 ml
DNase I		2 x 600 μl
RTL Lysis Buffer		80 ml
Buffer MCB		18 ml
96-Tip		1
Sample plate (DW Plate)	500μl Buffer MCB	1
Wash 1 Plate (DW Plate)	700μl Buffer MW130μl MagPure RNA Partilces	1
DNase Plate	Empty
Wash 2 Plate (DW Plate)	700μl Buffer MW1	1
Wash 3 Plate (DW Plate)	900μl Buffer MW2	1
Elution plate (DW Plate)	80μl RNase Free Water	1

Cat.No	Reagent	IVD3020-TL-06
Purification times		96 Preps
DNase I		2 x 600 μl
DNase Buffer		60 ml
RTL Lysis Buffer		60 ml
Buffer MCB		40 ml
96-Tip		12 PCS
2.0ml V-bottom plate	Row 1/7：500μl Buffer MCBRow 2/8：500μl Buffer MW1Row 3/9：emptyRow 4/10：30μl Magpure RNA Particles500μl Buffer MW2 Row 5/11：900μl Buffer MW2 Row 6/12：80μl RNase Free Water	6

Storage and Stability

MagPure RNA Particles should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, MagPure RNA Particles up to 8 weeks) at roomtemperature (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.

This product is suitable for rapid RNA extraction from tissue , cells, and other clinical samples. RNA can be used directly for RT-PCR, quantitative RT-PCR and so on.

What is Collagen?

Collagen is a fundamental component of the extracellular matrix, and the predominant protein in animals, constituting around 30% of total protein mass. A glycoprotein, it is well known for its triple helical structure. This is formed from three polypeptide α-chains with Gly-X-Y repeating residues (Gly for Glycine, X for proline, and Y for hydroxyproline).

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Types of Collagen

Over 28 types of collagens have been identified, with Type I collagen being the most abundant. It’s prevalent in ligaments, tendons, skin, and bone tissue. Its mature, insoluble form grants it remarkable strength, making it vital for the mobility of organisms. Collagen also has biochemical functions, influencing cell growth, proliferation, and differentiation.

This version of the kit is designed to detect and measure INSOLUBLE forms of collagen. Chose our Sircol 2.0 collagen kit if you need to analyse SOLUBLE collagen.

Applications of Collagen

Collagen, with its diverse properties, finds utility in various industries. It plays a role in medicine for wound healing and has an expanding role in tissue engineering and cell culture for biomedical purposes. It’s gaining popularity in the cosmetic industry for skin rejuvenation and is used in chemical formulations and the food industry as a functional food supplement and additive.

How does the Sircol assay detect collagen?

Sircol dye reagent contains Sirius Red – a linear anionic dye with sulphonic acid side chain groups. Under assay conditions the Sircol dye binds the basic groups of soluble collagen molecules. Maximal binding occurs in collagens possessing intact triple helix organisation as the highly ordered Gly-X-Yn helical structure of tropocollagen further contributes to dye binding. This results in a high degree of dye-collagen specificity. Affinity is progressively reduced during heat denaturation 4ºC due to the unwinding of the triple helix and formation of random chains.

‍Overview of the Sircol assay process:

Step 1. Samples being assayed for insoluble collagen must first undergo a 2-3 hour pre-treatment with Sircol Fragmentation reagent. This converts insoluble collagen into water-soluble gelatin can then be assayed.

Step 2. Addition of Sircol Dye Reagent to these pre-treated insoluble collagen samples results in the formation of a denatured collagen-dye complex. This complex then precipitates during the dye incubation period and is subsequently isolated by centrifugation, followed by washing to remove unbound dye. The Denatured collagen-bound dye is then  eluted and measured spectrophotometrically.

Step 3. The insoluble collagen content of unknown samples is quantified by comparison against a calibration curve prepared using a the denatured collagen standard supplied with the kit.

‍Assay range

100 – 1000 µg/ml

Limit of Detection

100µg/ml

Detection Method

Colorimetric Detection (556nm) (Endpoint)

Measurements per kit

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

Suitable Samples

The assay can be used to assess the rate of production of newly laid down collagen fibres during periods of rapid growth, development, tissue repair, remodeling and wound healing. Sources of material includes tissues, bone and calcified tissue.

*Insoluble collagens must be converted into soluble form prior to assay. Instructions and regents are provided with the kit., depending on sample this will require prior salt/acid/acid-pepsin extraction.

**non-mammalian collagens may result in a reduced limit of detection. We recommend use of an assay standard matched to the species under assay.‍

Many customers have found that the straightforward sample processing and analysis of Sircol make it a good alternative to conventional hydroxyproline analysis.

Precautions

This kit is designed for research use only. Not for use in diagnostic procedures.
Kit requires access to a centrifuge, water bath / heated block, as well as a spectrophotometer/colorimeter capable of absorbance detection at 556nm.
Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.

Sircol Insoluble Collagen kit contents:‍

1. Sircol Dye Reagent (1x110ml)

2. Denatured Collagen Reference Standard (1x5ml, 1.0mg/ml)

3. Acid-Salt Wash Reagent (1x20ml)

4. Fragmentation Reagent (1x110ml)

5. Alkali Reagent (1x110ml)

6. 2ml screw-cap tubes for preparation of samples.

7. Assay kit manual

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

As collagens mature, they become increasingly crosslinked and insoluble – characteristics necessary for key biophysical role that collagen plays in living organisms. Biocolor’s Sircol™ INSOLUBLE Collagen Kit is a dye-binding assay designed for accurate quantification and measurement such collagens. It is ideal for analyzing crosslinked / insoluble collagens from sources such as tissues, bone, and calcified tissue.

Propargyl-PEG7-alcohol is a propargyl linker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG7-alcohol is a propargyl linker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.