Propargyl-PEG4-Ms

Introducing the Fastin Assay Kit: Your Straightforward Solution for Elastin Quantification! Our user-friendly kit utilizes a dye-based method to measure elastin from in-vivo and in-vitro sources. It can be used to quantify various elastin forms, spanning from immature tropoelastin to mature, ‘insoluble’ elastin fibers.

Colorimetric Detection (513nm) (Endpoint)

Understanding Elastin: The Key to Tissue Flexibility

Tissues like lungs and arteries must maintain the ability to stretch and recoil repeatedly throughout an organism’s life. Elastin, a mature protein, is responsible for this elasticity and is usually present as insoluble fibers within the ECM. During development, these fibers are initially formed from a soluble precursor called tropoelastin.

‍What is the Fastin Assay?

The Biocolor Fastin assay is a user-friendly, dye-based means of quantifying elastins derived from both in-vivo and in-vitro sources. A variety of elastin forms can be assayed, from immature tropoelastin to mature ‘insoluble’ elastin fibres.

Further information on how the assay works can be found on the ‘Mode of Action‘ tab.

A list of suggested sample types can be found under the ‘Assay Specification‘ tab.

How does the Fastin assay detect Elastin?

The Fastin Dye Reagent contains an elastin-binding synthetic porphyrin, TPPS (5,10,15,20-tetraphenyl- 21H,23H-porphine). This affinity of TPPS for elastin was first observed when used as a ‘vital stain’ on live animals. Most tissues took up the dye initially but only elastin retained the TPPS molecules over time. [Winkelman, J. (1962), Cancer Res. 22, 589-596; Winkelman, J & Spicer, S. (1962), Stain Technol. 37, 303-305].

It has been proposed that the elastin binding of TPPS may be due to the retention of the acidic dye (which contains four charged sulfate groups) by the basic amino acid side chain residues of elastin.

‍How does the Fastin assay work?

Step 1. Incubation of samples containing soluble elastin with the Fastin Dye Reagent causes an elastin-dye complex to form. This insoluble complex then precipiates.

Step 2. Dye-labelled elastin is then isolated by centrifugation and the unbound dye removed. Elastin-bound dye is then eluted and measured spectrophotometrically.

Step 3. The elastin content of unknown samples can be calculated by comparison against a calibration curve prepared using a standard comprising water-soluble elastin (supplied with the kit).

Assay range

50 – 500µg/ml

Limit of Detection

50µg/ml

Detection Method

Colorimetric Detection (513nm) (Endpoint)

Measurements per kit

110 in total (allows a maximum of 48 samples to be run in duplicate alongside a standard curve).

Suitable Samples

In-vivo: tissues and fluids. Insoluble elastin will first require conversion to water soluble α-elastin using the oxalic acid reagents and extraction protocol supplied with the kit.  

In-vitro: Elastin produced by cells during 2D/3D cell culture. NB elastin in conditioned cell media is typically below the detection limit of the kit.

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Precautions

This kit is designed for research use only. Not for use in diagnostic procedures.
Kit requires access to a centrifuge, heated water bath or block, as well as a spectrophotometer or colorimeter capable of absorbance detection at 513nm.
Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.

Fastin elastin kit contents:‍

1. Dye Reagent (1x110ml)

2. α-elastin Reference Standard (1x5ml, 1.0 mg/ml soluble Bovine elastin)

3. Oxalic Acid (1x20ml) Precipitating Reagent (1x30ml)

4. Dye Dissociation Reagent (1x30ml)

5. Precipitating Reagent (1x30ml)

6. 1.5ml micro-centrifuge tubes for dye-labelling reaction.

7. Assay kit manual

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

Introducing the Fastin Assay Kit: Your Straightforward Solution for Elastin Quantification! Our user-friendly kit utilizes a dye-based method to measure elastin from in-vivo and in-vitro sources. It can be used to quantify various elastin forms, spanning from immature tropoelastin to mature, ‘insoluble’ elastin fibers.
Colorimetric Detection (513nm) (Endpoint)

t-Boc-aminooxy-PEG2-propargyl is a click chemistry crosslinker. The propargyl group is reactive with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry to yield a stable triazole linkage. t-Boc-aminooxy can be deprotected under mild acidic conditions. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

t-Boc-aminooxy-PEG2-propargyl is a click chemistry crosslinker. The propargyl group is reactive with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry to yield a stable triazole linkage. t-Boc-aminooxy can be deprotected under mild acidic conditions. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

The kit is developed for RNA quantification. The RNA Quantification HS kit includes HS Dye, HS Dilution Buffer, and two RNA Standards. Simply dilute the HS Dye with the HS Dilution Buffer, add RNA sample, then read the concentration using the Qubit Fluorometer. The assay is accurate for RNA concentrations from 250 pg/µL to 100 ng/µL (Figure 1). The RNA Quantification High Sensitivity Kit has several advantages over traditional RNA quantitation such as stability, sensitivity, and contaminant tolerance.

Features

• • • Optimized condition for using with the Qubit Fluorometer
    • Uses the Qubit RNA High Sensitivity assay setting
    • Cuts costs by 60%

The performance of the BioDynami RNA Quantification HS kit is nearly identical to that of Thermo Fisher’s Qubit RNA HS kit (figure below).

Common contaminants such as detergents, solvents, salts, free nucleotides, or protein are well tolerated in the assay (Table 1).

Qubit is a registered trademark of Thermo Fisher Scientific.

The kit is developed for RNA quantification. The RNA Quantification HS kit includes HS Dye, HS Dilution Buffer, and two RNA Standards. Simply dilute the HS Dye with the HS Dilution Buffer, add RNA sample, then read the concentration using the Qubit Fluorometer. The assay is accurate for RNA concentrations from 250 pg/µL to 100 ng/µL (Figure 1). The RNA Quantification High Sensitivity Kit has several advantages over traditional RNA quantitation such as stability, sensitivity, and contaminant tolerance.