Isolate high quality total RNA (including small RNA and microRNA) and all sizes of circulating and exosomal RNA, including microRNA from urine and cerebrospinal fluid (CSF) samples
Small urine and CSF input ranging from as low as 0.5 mL to 1 mL
No phenol extractions
Very sensitive and linear down to a few cells without the need for carrier RNA
Bind and elute all RNA irrespective of size or GC content, without bias
Rapid and convenient spin column protocol
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
Norgen’s Urine microRNA Purification Kit provides a rapid method for the isolation and purification of small RNA molecules (All sizes, including < 200 nt) from urine samples. These small RNAs include regulatory RNA molecules such as microRNA (miRNA) as well as tRNA and 5S rRNA. Small RNA molecules are often studied due to their ability to regulate gene expression. Typically miRNAs are 20-25 nucleotides long and regulate gene expression by binding to mRNA molecules and affecting their stability or translation. Several recent studies have shown that miRNA regulates cell growth and apoptosis. Furthermore, clinical and experimental analyses suggested that miRNAs may function as a novel class of oncogenes or tumor suppressor genes. MicroRNA expression profiles of different tumor types, relative to their normal tissues, have recently been shown to provide phenotypic signatures for particular cancer types. Unique patterns of aberrant miRNA expression may serve as molecular biomarkers for tumor diagnosis, prognosis of disease-specific outcomes, and prediction of therapeutic responses.
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
Component
Cat. 29000 (25 preps)
Lysis Buffer A
2 x 20 mL
Wash Solution A
18 mL
Elution Solution A
6 mL
Mini Spin Columns
25
Collection Tubes
25
Elution Tubes (1.7 mL)
25
Product Insert
1
Other Products
R6628 MagPure Serum miRNA Kit
Product Info
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Product Info
Introduction
The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Details
Specifications
Features
Specifications
Main Functions
Isolation miRNA from 0.3-0.5ml serum or plasma using magnetic particles
Applications
RT-PCR, cDNA synthesis, second generation sequencing
Purification method
Polydisperse magnetic beads
Purification technology
Magnetic beads technology
Process method
Manual or automatic
Adaptive instrument
Nucleic acid extractor, pipetting workstation
Sample type
Serum, plasma, acellular samples
Sample amount
300μl
Principle
This product is based on the purification method of high binding magnetic particles. The sample material is denatured in Lysis Buffer. The protein is then precipitated by Protein Precipitation Solution and pelleted by centrifugation. After adding magnetic particles and binding solution, RNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption.The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally RNA was eluted by Elution Buffer.
Advantages
Safe – no phenol chloroform extraction and no use of Trizol reagent
Fast – several samples can be extracted in 60 minutes by column method
Kit Contents
Contents
R662801
R662802
R662803
Purification Times
48 Preps
96 Preps
5 x 96 Preps
MagPure Particles N
1.7 ml
3.5 ml
17 ml
Buffer CFL
6 ml
12 ml
60 ml
Buffer CPL
1.8 ml
3.5 ml
20 ml
Buffer MGW1*
30 ml
60 ml
250 ml
Buffer MW2*
12 ml
25 ml
100 ml
RNase Free Water
10 ml
20 ml
60 ml
Storage and Stability
MagPure Particle N should be stored at 2–8°C upon arrival. However, short-term storage (up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
Document
The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Collagen is a fundamental component of the extracellular matrix, and the predominant protein in animals, constituting around 30% of total protein mass. A glycoprotein, it is well known for its triple helical structure. This is formed from three polypeptide α-chains with Gly-X-Y repeating residues (Gly for Glycine, X for proline, and Y for hydroxyproline).
Types of Collagen
Over 28 types of collagens have been identified, with Type I collagen being the most abundant. It’s prevalent in ligaments, tendons, skin, and bone tissue. Its mature, insoluble form grants it remarkable strength, making it vital for the mobility of organisms. Collagen also has biochemical functions, influencing cell growth, proliferation, and differentiation.
This version of the kit is designed to detect and measure INSOLUBLE forms of collagen. Chose our Sircol 2.0 collagen kit if you need to analyse SOLUBLE collagen.
Applications of Collagen
Collagen, with its diverse properties, finds utility in various industries. It plays a role in medicine for wound healing and has an expanding role in tissue engineering and cell culture for biomedical purposes. It’s gaining popularity in the cosmetic industry for skin rejuvenation and is used in chemical formulations and the food industry as a functional food supplement and additive.
How does the Sircol assay detect collagen?
Sircol dye reagent contains Sirius Red – a linear anionic dye with sulphonic acid side chain groups. Under assay conditions the Sircol dye binds the basic groups of soluble collagen molecules. Maximal binding occurs in collagens possessing intact triple helix organisation as the highly ordered Gly-X-Yn helical structure of tropocollagen further contributes to dye binding. This results in a high degree of dye-collagen specificity. Affinity is progressively reduced during heat denaturation 4ºC due to the unwinding of the triple helix and formation of random chains.
Overview of the Sircol assay process:
Step 1. Samples being assayed for insoluble collagen must first undergo a 2-3 hour pre-treatment with Sircol Fragmentation reagent. This converts insoluble collagen into water-soluble gelatin can then be assayed.
Step 2. Addition of Sircol Dye Reagent to these pre-treated insoluble collagen samples results in the formation of a denatured collagen-dye complex. This complex then precipitates during the dye incubation period and is subsequently isolated by centrifugation, followed by washing to remove unbound dye. The Denatured collagen-bound dye is then eluted and measured spectrophotometrically.
Step 3. The insoluble collagen content of unknown samples is quantified by comparison against a calibration curve prepared using a the denatured collagen standard supplied with the kit.
Assay range
100 – 1000 µg/ml
Limit of Detection
100µg/ml
Detection Method
Colorimetric Detection (556nm) (Endpoint)
Measurements per kit
110 in total (allows a maximum of 46 samples to be run in duplicate alongside a standard curve).
Suitable Samples
The assay can be used to assess the rate of production of newly laid down collagen fibres during periods of rapid growth, development, tissue repair, remodeling and wound healing. Sources of material includes tissues, bone and calcified tissue.
*Insoluble collagens must be converted into soluble form prior to assay. Instructions and regents are provided with the kit., depending on sample this will require prior salt/acid/acid-pepsin extraction.
**non-mammalian collagens may result in a reduced limit of detection. We recommend use of an assay standard matched to the species under assay.
Many customers have found that the straightforward sample processing and analysis of Sircol make it a good alternative to conventional hydroxyproline analysis.
Precautions
This kit is designed for research use only. Not for use in diagnostic procedures. Kit requires access to a centrifuge, water bath / heated block, as well as a spectrophotometer/colorimeter capable of absorbance detection at 556nm. Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.
Sircol Insoluble Collagen kit contents:
1. Sircol Dye Reagent (1x110ml)
2. Denatured Collagen Reference Standard (1x5ml, 1.0mg/ml)
3. Acid-Salt Wash Reagent (1x20ml)
4. Fragmentation Reagent (1x110ml)
5. Alkali Reagent (1x110ml)
6. 2ml screw-cap tubes for preparation of samples.
7. Assay kit manual
NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details.
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As collagens mature, they become increasingly crosslinked and insoluble – characteristics necessary for key biophysical role that collagen plays in living organisms. Biocolor’s Sircol™ INSOLUBLE Collagen Kit is a dye-binding assay designed for accurate quantification and measurement such collagens. It is ideal for analyzing crosslinked / insoluble collagens from sources such as tissues, bone, and calcified tissue.
Attogene’s Montmorillonite Bentonite Clay binds toxins with high affinity and high capacity. Montmorillonite has been shown to be the active ingredient in bentonites. The proposed mechanism of action of Attogene’s Montmorillonite Bentonite Clay is through the adsorption of toxins (mainly onto the interlayer spaces of montmorillonite). Attogene’s Montmorillonite Bentonite Clay is a common anti-caking agent in animal feeds to adsorb aflatoxins and diminish their bioavailability. Its composite for application in drug system can be formed through the utilization of anionic, cationic, and nonionic surfactants.
Bentonite is a promising rock of clay which is found in nature. It is a major source of montmorillonite in nature. It is a rock produced of highly colloidal and plastic clays mainly comprised of montmorillonite. In addition to montmorillonite, bentonite may compose of some amount of crystalline quarz, cristobalite, and feldspar. Montmorillonite nanoclay is used as a drug carrier system and as an additive. Its composite for application in drug system can be formed through the utilization of anionic, cationic, and nonionic surfactants. These are used to improve basal spacing resulting in organo clay to be utilised in drug loading and drug release.
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Promotes flocculation of cyanobacteria cell debris, binds residual phosphate compounds and promotes gradual settling of the cyanobacterial biomass on the sediment.
