t-Boc-aminooxy-PEG5-propargyl

Overview

• CE-IVDR marked in accordance with the European Commission Regulation (EU) No. 2017/746.
• Ideal for use in in vitro diagnostic workflows
• Sample collection is non-invasive and painless
• Fast and easy processing using a rapid spin-column format
• Isolate high quality genomic DNA to be used in downstream in vitro diagnostic applications 
• Compatible with preserved saliva samples collected using Norgen’s Saliva DNA Collection and Preservation Devices, as well as fresh saliva samples 

This kit provides a fast and simple spin column procedure for isolating genomic DNA from saliva samples collected and preserved using Norgen’s Saliva DNA Collection and Preservation Devices (Cat. 49000), as well as fresh saliva samples.

Saliva DNA purified using Norgen’s Saliva DNA Isolation Kit Dx kit is of the highest quality, and can be used with any downstream in vitro diagnostic application employing enzymatic amplification or other enzymatic modifications of DNA followed by signal detection or amplification.

Background

Saliva represents an excellent non-invasive alternative to blood collection.  Human genomic DNA extracted from buccal epithelial cells and white blood cells found in saliva can be used in various applications in diagnostics. Saliva DNA can be used for the detection of biomarkers to diagnose a disease, follow the diseases progress or monitor the effects of a particular treatment. Saliva DNA can also be used to diagnose particular types of infections. Isolation of DNA from saliva has become an attractive alternative to isolation from blood or tissue due to the fact that sample collection is non-invasive, the samples can be collected by individuals with little training, and no special equipment is required. Norgen’s Saliva DNA Isolation Kit provides a fast and simple procedure for isolating genomic DNA from both preserved saliva samples and fresh saliva samples.

NOTE: This product is not available for sale in the United States.

Details

Supporting Data

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Kit Specifications
Maximum Saliva Input	0.5 mL preserved saliva 0.25 mL fresh saliva
Average Yield from 0.25 mL of Saliva*	3 – 7 μg
Average Purity (OD260/280)	1.7 – 2.1
Time to Complete 10 Purifications	30 minutes

* Average yield will depending on the donor

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. All solutions and plastics can be used until the expiration date specified on their labels. The Proteinase K can be stored at either room temperature or 4^oC.

Component	Cat. Dx45400 (50 preps)
Lysis Solution	30 mL
Proteinase K in Storage Buffer	1.2 mL
Binding Solution	12 mL
Wash Solution	18 mL
Elution Buffer	12 mL
Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Hydroxy-PEG3-DBCO is a PEG linker containing a DBCO moiety and a terminal primary hydroxyl group. The hydroxyl can react with a variety of functional groups and the hydrophilic PEG spacer arm can provide better solubility to labeled molecules. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Hydroxy-PEG3-DBCO is a PEG linker containing a DBCO moiety and a terminal primary hydroxyl group. The hydroxyl can react with a variety of functional groups and the hydrophilic PEG spacer arm can provide better solubility to labeled molecules. DBCO is commonly used for copper-free Click Chemistry reactions. 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.