Principle: Peptone provide carbon and nitrogen sources to meet the needs of bacterial growth; lactose are fermentable sugars; selenite, sodium hydrogen inhibit Gram-positive bacteria and gram-negative enterobacteria most non-Salmonella; phosphate-buffered agent; L- cystine as a reducing agent.
How to use: 1. Weigh 23g of the product , adding 1 L of distilled or deionized water , heated to boiling stirring until completely dissolved, dispensing flask, cooled to room temperature . 2. Pipette 10mL of pre-enrichment sample broth or 25mL and transferred species in the liquid sample flask in a sterile environment. 3.Place into incubator, cultured at 36 ± 1 for 18-24h. 4. Observe the results.
Quality control: Quality control strains were inoculated ,and cultured at 36 ± 1 for 18-24h ,results show as follows: strain name strain code growth feature Salmonella typhi CMCC (B) 50071 good red, cloudy Salmonella typhimurium CMCC (B) 50115 good red, cloudy Escherichia coli ATCC25922 — remain unchanged
Storage: Store in a dark, cool and dry place, tighten the cap immediately after use. Storage period of three years.
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
Ideal magnet for all of your low volume needs. From Low elution 96-well PCR applications to 384-well, the X396 has you covered. No need to purchase two separate plates anymore. 96-well volumes as low as 5 µL are achieved from 96-well, PCR plates on the inside of the magnet, while the outside of each magnet will pull beads to well sides in 384-well plates.
ANSI/ SBS Footprint (127.75mm x 85.50mm) to fit into any automated liquid handling robot on bottom. ANSI/SBS footprint on top to accept all common microplates.
Integrated Cushion base for maximum recovery. Helps aid in set-up, robot positioning inconsistencies, and labware consumable inconsistencies
Features include solid aluminum alloy construction and hard coat anodized finish for years of trouble-free use, and compatible with any magnetic beads
Ideal magnet for all of your low volume needs. From Low elution 96-well PCR applications to 384-well, the X396 has you covered. No need to purchase two separate plates anymore. 96-well volumes as low as 5 µL are achieved from 96-well, PCR plates on the inside of the magnet, while the outside of each magnet will pull beads to well sides in 384-well plates.
