DBCO-PEG8-acid is an analog of DBCO-Acid with hydrophilic PEG linker and a DBCO group. The DBCO groups is commonly used for Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-PEG8-acid is an analog of DBCO-Acid with hydrophilic PEG linker and a DBCO group. The DBCO groups is commonly used for Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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.

Introduction

This product is suitable for rapid extraction of DNA from FFPE sample, tissue, cells, blood, swabs, blood spots, semen and other clinical samples. This product uses size selection magnetic beads, which can selectively remove small sizes of DNA (100-300bp) from FFPE samples by adjusting the amount of binding solution, so as to improve the effective data volume of downstream NGS.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from FFPE using high bind beads
Applications	RT-PCR, northern blot, poly A purification, nucleic acid protection and in vitro translation, etc.
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Large quantities of solids
Sample amount	Appropriate
Elution volume	≥50μl
Time per run	30 – 120 minutes

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 Protease. 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

1.Use non-toxic dewaxing solution without contact with xylene

2.Efficient removal of formaldehyde modification on DNA and improvement of PCR sensitivity

3. One of the best FFPE DNA extraction kits on the market, the same effect of paraffin slice extraction as top brand, and the effect of puncture sample extraction is even better than top brands

Kit Contents

Contents	D632301B	D632302B
Purification Times	48 Preps	96 Preps
MagBind Particles	1.1 ml	2 x 1.1 ml
RNase A	10 mg	20 mg
Proteinase K	24 mg	48 mg
Protease Dissolve Buffer	3 ml	6 ml
Buffer DPS	60 ml	100 ml
Buffer ATL	15 ml	30 ml
Buffer AL	15 ml	30 ml
Buffer BD*	6 ml	15 ml
Buffer BXW1*	13 ml	44 ml
Elution Buffer	15 ml	30 ml

Storage and Stability

RNase A, Proteinase K and MagBind Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 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

This product is suitable for rapid extraction of DNA from FFPE sample, tissue, cells, blood, swabs, blood spots, semen and other clinical samples. This product uses size selection magnetic beads, which can selectively remove small sizes of DNA (100-300bp) from FFPE samples by adjusting the amount of binding solution, so as to improve the effective data volume of downstream NGS.