t-Boc-N-Amido-PEG3-propargyl enables formation of triazole linkage with azide-bearing compound via copper catalyzed Click Chemistry. Under mild acidic conditions, t-Boc group can be removed to yield the free amine. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Detail
t-Boc-N-Amido-PEG3-propargyl enables formation of triazole linkage with azide-bearing compound via copper catalyzed Click Chemistry. Under mild acidic conditions, t-Boc group can be removed to yield the free amine. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Our Blyscan™ Glycosaminoglycan Kit has been a ‘go-to’ Solution for reliable sGAG and Proteoglycan Analysis for many years! Blyscan utilises a dye-binding approach to quantitatively measure sulfated glycosaminoglycans (sGAG) and proteoglycans in cells, tissues and fluids from a wide range of in-vivo and in-vitro sources.
Colorimetric Detection (656nm) (Endpoint)
Understanding Glycosaminoglycans (GAGs) and Proteoglycans
Glycosaminoglycans (GAGs) are a type of negatively charged polysaccharide that play crucial roles in various biological processes. They are composed of repeated disaccharide units, typically of N-acetylated or N-sulfated hexosamine paired with a uronic acid (GlcA or IdoA) or galactose. Sulfate groups can also be added to give sulfated GAGs an overall negative charge that influences cell interactions and also enable binding by our Blyscan dye reagent.
Common examples of GAGs include Chondroitin Sulfate, Dermatan Sulfate, Heparin, Heparan Sulfate, and Keratan Sulfate. Note that Hyaluronic Acid is a non-sulfated GAG and cannot be detected by the Blyscan assay. If you need to measure hyaluronic acid instead, we recommend using our Purple-Jelley kit!
The Role of Glycosaminoglycans in Tissues
GAGs and proteoglycans have essential functions in tissues and organisms, providing biophysical support through scaffolding and maintaining cartilage hydration. They also play a vital role in biochemical processes such as cell adhesion and signalling.
What is the origin of the Blyscan assay name?
Blyscan is an Old English word meaning ‘to shine’ and from which the word ‘blush’, (blushing), may have been derived. This was an appropriate choice as the Blyscan Assay contains a blue dye which ‘blushes’ bright pink when it binds to sulphated glycosaminoglycans!
How does the Blyscan assay work?
Step 1. Blyscan dye reagent contains DMMB dye in an optimised buffer. Addition of Dye reagent to samples containing sGAG results in the formation of a dye/sGAG complex due to a charge interaction between dye and GAG sulfate groups.
Step 2. Over a 30 minute incubation Dye-labelled sGAGs precipitate out of solution and are collected by centrifugation. Following removal of unbound dye, the remaining bound dye is released from the complex by addition of dye dissociation reagent. Released dye is quantified spectrophotometrically.
Step 3. The sGAG content of unknown samples may be quantified by comparison against a calibration curve prepared using a standard of purified Chondroitin-4-sulfate supplied with the kit.
A list of suggested sample types can be found under the ‘Assay Specification‘ tab.
The Blyscan Dye reagent is formulated to miminise binding to other charged sample components such as nucleic acids, a problem with some older dye-based sGAG assays.
Assay range
2.5 – 50µg/ml
Limit of Detection
2.5µg/ml
Detection Method
Colorimetric Detection (656nm) (Endpoint)
Measurements per kit
110 in total (allows a maximum of 48 samples to be run in duplicate alongside a standard curve).
In-vivo: Liquid samples, including fluids such as urine, amniotic or synovial fluid.
In-vitro: Solid samples, such as deposited ECM on 2D/3D culture surfaces.by enzymatic treatment
In-vivo: Liquid samples, Culture media during 2D/3D cell culture.
The assay requires that sulfated polysaccahrides or sGAGs are in a soluble form. A preliminary enzymatic extraction step is required for solid samples (enzyme not supplied with kit).
The assay is not suitable for use with samples containing alginates or that comprise degraded sulfated disaccharide fragments.
Precautions
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 absorbance detection at 656nm. Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.
Blyscan sGAG kit contents:
1. Blyscan Dye Reagent (1x110ml)
2.sGAG Reference Standard (1x5ml, 100µg/ml Bovine tracheal chondroitin 4-sulfate)
3. Dissociation Reagent (1x110ml)
4. Sodium Nitrite (1x15ml)
5. Acetic Acid (1x15ml)
6. Ammonium Sulfamate (1x15ml)
7. 1.5ml micro-centrifuge tubes for dye-labelling reaction.
8. Assay kit manual
NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details.
Document
Our Blyscan™ Glycosaminoglycan Kit has been a ‘go-to’ Solution for reliable sGAG and Proteoglycan Analysis for many years! Blyscan utilises a dye-binding approach to quantitatively measure sulfated glycosaminoglycans (sGAG) and proteoglycans in cells, tissues and fluids from a wide range of in-vivo and in-vitro sources. Colorimetric Detection (656nm) (Endpoin
The Norgen PCRSizer 100 bp DNA Ladder is prepared to ensure quality and batch-to-batch consistency. Our PCRSizer contains ten discrete fragments ranging from 100 bp to 1000 bp in 100 bp increments with a higher intensity reference band at 500 bp. This Ladder is ideal for PCR product size confirmation.
Contents: 1mL of premixed DNA ladder (0.5µg/10µL) in loading buffer (10mM EDTA, 10% glycerol, 0.015% bromophenol blue, and 0.17% SDS).
PCR Sizer 100 bp DNA Ladder (Cat# 11400) – 100 loads
Ladder Properties: • Ten discrete bands, ranging from 100 bp to 1,000 bp • Higher intensity band at 500 bp for easy reference
Fragment
Size (bp)
Mass (ng)
1
1000
78
2
900
70
3
800
62
4
700
55
5
600
47
6
500
78
7
400
31
8
300
24
9
200
31
10
100
24
Recommended Use:
Mix thoroughly. For best results, load 10µL of DNA ladder per well. For precise mass determination with a densitometer, stain gel after electrophoresis using 0.5µg/mL ethidium bromide for 30-40 minutes. The table above shows the size and mass for each band based on 10µL ladder per well.
Storage:
Stable at room temperature. For longer term storage, -20°C is recommended.
This ladder was standardized using 10µL of DNA per lane on a 0.8 cm thick, 13 x 15 cm, 1.0% agarose gel run in TAE buffer.