Description

Specifications

Clone	IHC125
Source	Mouse Monoclonal
Positive Control	Ovarian Serous Carcinoma
Dilution Range	1:200

Annexin A1 (ANXA1) is a membrane protein that plays a role in innate and adaptive immunity by controlling the biosynthesis of inflammation, prostaglandins, and leukotriene mediators. This target is overexpressed in 97% of all samples from patients with with hairy cell leukemia, and is absent in other B-cell lymphomas. High ANXA1 expression is frequently associated with advanced stage esophageal and esophagogastric junction adenocarcinoma, and is also linked to advanced and metastatic disease states.

Overview

• Extract high quality & quantity total RNA including miRNA 
• No phenol step required; isolate all RNA in one fraction
• Genomic DNA Removal Column for efficient elimination of gDNA
• Bind & elute all RNA irrespective of size or GC content, without bias
• Efficiently extract small RNA irrespective of GC content
• Very sensitive & linear down to a few cells without the need for carrier RNA
• Convenient & fast spin column format
• Isolate from a wide variety of specimens
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit purifies total RNA including miRNA from biological samples and employs an extra column for the efficient removal of contaminating gDNA, thereby replacing the enzymantic DNase step. 

Efficiently extract total RNA from a range of samples including cells, bacteria, yeast, virus and bodily fluids including plasma/serum, blood, saliva, CSF and more.  Extract high quality and purity RNA with excellent RIN values and A260/A280 suitable for downstream applications including qRT-PCR, RT-PCR, microarrays, NGS and more.

The kit purifies all sizes of RNA from large mRNA, lncRNA down to microRNA (miRNA) in the same fraction without the requirement of phenol.  Isolate all RNA sequences at an equal rate irrespective of size. Moreover, when the RNA sequences are small (e.g. miRNA), the column binds small RNAs regardless of their GC content.

Details

Supporting Data

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Kit Specifications
Maximum Binding Capacity Per Column	Up to 50 μg RNA
Maximum Loading Volume Per Spin Column	650 μL
Size of RNA Purified	All sizes, including < 200 nt
Time to Complete 10 Purifications	25 minutes
Maximum Amount of Starting Material
Liver Tissue	20 mg
Heart Tissue	5 mg
Kidney Tissue	20 mg
Brain Tissue	25 mg
Lung Tissue	20 mg
Spleen Tissue	20 mg
Whole Blood	100 µL
Hela Cells	3 x 106
CHO	3 x 106
Yeast	1 x 108 cells
E.coli	1 x 109 CFU/mL
Bacteria	1 x 109 cells
Plasma/Serum	200 µL
Fungi	50 mg
Plant Tissue	50 mg
Viruses	≤100 µL of viral suspension
Average Yield
HeLa Cells (1 x 106 cells)E. coli (1 x 109 cells)15 µg50 µg

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 48300 (50 preps)	Cat. 48400 (100 preps)
Buffer RL	40 mL	2 x 40 mL
Wash Solution A	38 mL	2 x 38 mL
Elution Solution A	6 mL	2 x 6 mL
RNA Purification Columns	50	100
Genomic DNA Removal Columns	50	100
Collection Tubes	100	200
Elution Tubes (1.7 mL)	50	100
Product Insert	1	1

Introduction

The HiPure Plasmid DNA Mega Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 10 mg high copy number plasmid DNA can be purified from 500 mL overnight culture.

Details

Specifications

Features	Specifications
Main Functions	Isolation up to 10mg endotoxin-free plasmid DNA from 500ml bacterial culture
Applications	Cell transfection, animal injection, etc.
Purification method	Mega spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	High copy plasmid vector
Sample amount	500ml LB
Yield	1~10mg
Elution volume	≥2ml
Time per run	≤70 minutes
Liquid carrying volume per column	50ml
Binding yield of column	10mg

Principle

The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
• Fast – silica gel column purification is much faster than ion exchange
• High yield – up to 10mg plasmid can be binded in one column
• Low endotoxin – the obtainedplasmid can be directly used for cell transfection and animal injection, etc.

Kit Contents

Contents	P111602	P111603
Purification Times	10 Preps	50 Preps
RNase A	60 mg	2 x 150 mg
Buffer E1	220 ml	2 x 550 ml
Buffer E2	220 ml	2 x 550 ml
Buffer E3	220 ml	2 x 550 ml
Buffer E4	220 ml	2 x 550 ml
Buffer E5	120 ml	550 ml
Buffer PW2	25 ml	2 x 100 ml
Elution Buffer	30 ml	120 ml
HiPure EF Maxi Columns	10	50
Lysate Clear Maxi Syringe	10	50
50 ml Collection Tubes	20	100

Storage and Stability

The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve.

Purchase Guide

Name	CAT NO	Features	Sampleamount	Elutionvolume	Yield
HiPure Plasmid Mini Kit	P1001	Classic, Regular application	1-5ml LB	30-50μl	5–35µg/5ml
HiPure Plasmid Mini Kit II	P1002	Classic, Regular application	5-15ml LB	75-100μl	20-80µg/15ml
HiPure Plasmid Plus 96 Kit	P1006	Classic, Regular application	1-1.5ml LB	70-150μl	1-15μg/1ml
HiPure Fastfilter Plasmid Midi Kit	P1013	Fast	30-50ml LB	0.3-0.8ml	50–250µg/50ml
HiPure Fastfilter Plasmid Maxi Kit	P1014	Fast	100-200ml LB	0.5-1.5ml	0.4–1mg/200ml
Low endotoxin/Endotoxin-free Plasmid
HiPure Plasmid EF Maxi Kit	P1156	Classic method, General for all kinds of vectors	100-200ml LB	≥0.7ml	200–1500µg
HiPure Plasmid EF Mini Kit	P1154	Recommend for low copy vector, Thoroughly remove RNA	5-15ml LB	≥60μl	10–80µg
HiPure Plasmid EF 96 Kit	P1157	Recommend for low copy vector, Thoroughly remove RNA	1-5ml LBx96	≥75μl	30µg
HiPure Plasmid EF Midi Kit	P1231	Recommend for High copy vector, Low elution volume, High concentration	25-50ml LB	≥100μl	10-500μg
HiPure Plasmid EF Mega Kit	P1116	Recommend for High copy vector(>3μg/ml)	500ml LB	≥2ml	1-10mg

For any technical problems or customized products, please contact us.

F&Q about Endotoxin-free Plasmid Extraction Kit — P1156 ←click here

The HiPure Plasmid DNA Mega Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 10 mg high copy number plasmid DNA can be purified from 500 mL overnight culture.