Description

Clone	IHC506
Source/Clonality	Mouse Monoclonal
Positive Control	Appendix, Uterus, Vessel Wall
Dilution Range	1:200

Actin is part of the cytoskeletal system of all cell types. Smooth Muscle Actin is found in myofibroblasts and myoepithelium, but not in cardiac and skeletal muscles. Labelling of smooth muscle actin in concert with muscle specific actin staining can allow for differentiation between rhabdomyosarcoma and leiomyosarcoma, as Muscle specific actin is found in rhabdomyoblasts, while smooth muscle actin is found in leiomyosarcomas.

Blyscan™ – sulfated Glycosaminoglycan (sGAG) assay kit

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).

Suitable Samples

In-vivo: Solid samples, including cartilage, bone, connective tissue, tumour tissue

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.

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

• 100 Lateral Flow Dipsticks (4.5mm)
• 20 mL Assay running buffer
• 100 wells with support plate
• Controls

Description

Attogene Universal Lateral Flow Assay Kits are a convenient ready-to-use kit for quick and cost-effective development of a lateral flow dipstick assay for detection of  DNA and RNA products.

Formats (strep gold conjugate pad):

Detection of nucleic Acid (DNA or RNA) requires the use of a biotin and FAM/FITC-labelled primer during amplification.  Test line: anti-biotin, Control Line: GAM

Multiplex detection of nucleic Acid (DNA or RNA) requires the use of a biotin, FITC/FITC and Dig labelled primers during amplification.: Test Line #1: anti FITC/FAM, Line #2: anti-Dig, Line #3 GAM.

Inquire about custom configurations: sales@attogene.com

Kit Components 

• 100 -4.5mm Lateral Flow Dipsticks
• 20 mL Sample assay running buffer

Features & Benefits

• Can be used for development of a lateral flow assay for detection of a variety of different molecules such as amplified DNA products from PCR, LAMP and RPA reactions.
• No need to stripe capture antibodies
• No expensive equipment required
• Cost-effective way to screen for further downstream lateral flow assay development.

Attogene Universal Lateral Flow Assay Kits are a convenient ready-to-use kit for quick and cost-effective development of a lateral flow dipstick assay for detection of  DNA and RNA products.

Formats (strep gold conjugate pad):

Detection of nucleic Acid (DNA or RNA) requires the use of a biotin and FAM-labelled primer during amplification.  Test line: anti-biotin, Control Line: GAM

Multiplex detection of nucleic Acid (DNA or RNA) requires the use of a biotin, FITC and Dig labelled primers during amplification.: Test Line #1: anti FITC/FAM, Line #2: anti-Dig, Line #3 Biotin.

Inquire about custom configurations: sales@attogene.com

Kit Components 

• 100 -4.5mm Lateral Flow Dipsticks – widest dipsticks around!
• 20 mL Sample assay running buffer

Features & Benefits

• Can be used for development of a lateral flow assay for detection of a variety of different molecules such as amplified DNA products from PCR, LAMP and RPA reactions.
• No need to stripe capture antibodies
• No expensive equipment required
• Cost-effective way to screen for further downstream lateral flow assay development.

100 Lateral Flow Dipsticks (4.5mm)
20 mL Assay running buffer
100 wells with support plate
Controls