Introduction

Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from bacteria, yeast cells
Applications	RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Bacteria, yeast cells
Sample amount	Bacteria: <10^9；Yeast cells：<2 x10^7
Elution volume	≥30μl
Time per run	≤40 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

Advantages

• Fast – several samples can be extracted in 30 minutes
• High purity – the purified RNA can be directly used in various downstream applications
• High recovery – RNA can be recovered at the level of PG
• Good repeatability – silica gel column purification technology can obtain ideal results every time
• Broad spectrum – it can deal with various bacteria, including Gram-positive bacteria that are difficult to be lysed
• Sufficient components – the kit contains carried wall breaking enzymes and glass beads

Kit Contents

Contents	R418202	R418203
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	250
Glass Beads (0.1-0.6mm)	30 g	150 g
DNase I	600 μl	5 x 600 μl
DNase Buffer	6 ml	30 ml
Protease Dissolve Buffer	1.8 ml	15 ml
Buffer ATL	50 ml	200 ml
Buffer PHC	50 ml	200 ml
Buffer GXP2*	20 ml	100 ml
Buffer RW1	50 ml	250 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

Buffer PHC should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Buffer PHC up to 12 weeks) at room temperature(15–25°C) does not affect their performance. The remaining kit components can be stored at roomtemperature (15–25°C) and are stable for at least 18 months under these conditions.

Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.

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.

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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, 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

Sulfo DBCO-PEG4-Maleimide is a water-soluble click chemistry reagent with a maleimide group and a DBCO moiety. The hydrophilic PEG spacer arm improves solubility in aqueous buffers. Maleimide group specifically and efficiently reacts with thiols to form stable thioether bonds. The low mass weight will add minimal spacer to modified molecules and will enable simple and efficient incorporation of DBCO moiety onto cysteine-containing peptides or other thiol-containing biomolecules. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Sulfo DBCO-PEG4-Maleimide is a water-soluble click chemistry reagent with a maleimide group and a DBCO moiety. The hydrophilic PEG spacer arm improves solubility in aqueous buffers. Maleimide group specifically and efficiently reacts with thiols to form stable thioether bonds. The low mass weight will add minimal spacer to modified molecules and will enable simple and efficient incorporation of DBCO moiety onto cysteine-containing peptides or other thiol-containing biomolecules. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.