NEST 15 mL Centrifuge Tube (500 count)

Description

Attogene’s Carbon Dioxide Enzymatic Assay Kit is a simple, direct method for measuring Carbon Dioxide levels in the environment. The assay uses a coupled enzyme assay to detect CO2 (as HCO3-) as follows. In the first step, the bicarbonate condenses with phosphoenol pyruvate to form oxalate (and phosphoric acid); this reaction is catalyzed by the enzyme Phosphoenolpyruvate Decarboxylase, PEPC. The oxalate is then enzymatically reduced by the enzyme Malate Dehydrogenase (using an NADH cofactor) to form malate and NAD+.

Overview

• Isolation of exosomal RNA molecules from urine samples
• Rapid and convenient spin-column protocol
• Isolate inhibitor-free urinary microRNA for most downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Optional protocol for purification of exosomal proteins for western blot analysis

This kit provides a rapid spin column procedure for the isolation of exosomal RNA from urine samples. Users can simultaneously concentrate and isolate high quality exosomal RNA, including microRNA, for use in sensitive downstream assays such as RT-PCR, qRT-PCR, NGS, microarrays and more. The protocol can be completed in under 50 minutes. Urine volumes of 1 to 10 mL can be processed easily and rapidly. All sizes of RNA are recovered at an equal rate without the need for using hazardous chemicals like phenol.

Background

Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascite fluids, among other biological fluids. These vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which can inform about the cell type from which the exosomes are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNases they can be efficiently recovered from biological fluids, such as urine.

Details

Supporting Data

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Kit Specifications
Minimum Urine Input	1 mL
Maximum Urine Input	10 mL
Size of RNA Purified	Small exosomal RNA species
Time to Complete Purification	~ 50 minutes

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Component	Cat. 47200 (50 preps)
Slurry B1	18 mL
Binding Buffer A	20 mL
Lysis Buffer A	2 x 20 mL
Wash Solution A	38 mL
Elution Solution A	6 mL
Mini Filter Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Propargyl-PEG4-bromide is a crosslinker that can be used in copper catalyzed azide-alkyne Click Chemistry to form a stable triazole linkage with azides. The bromide (Br) acts as a good leaving group for nucleophilic substitution reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG4-bromide is a crosslinker that can be used in copper catalyzed azide-alkyne Click Chemistry to form a stable triazole linkage with azides. The bromide (Br) acts as a good leaving group for nucleophilic substitution reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.