TECHNICAL RESOURCE

HRP-SA and TrueBlue peroxi- dase substrate: Orcein and. Eosin Y counterstains. Southern and Northern blotting; mRNA in situ hybridization. Detection of single.

HiPer® Southern Blotting Teaching Kit

Southern blotting or Southern hybridization is a widely used technique in peroxidase (HRP)-conjugated streptavidin which attaches to the hybridized DNA.

TECHNICAL RESOURCE - Strip Northern and Southern Blots

Following exposure of a Northern or Southern blot (either by classical 17095) test for stripping efficiency by re-applying streptavidin-HRP and add ...

Untitled

solanacearum hrp genes and hrp gene clusters of P. syringae pv. formed Southern blot hybridizations between the three. P. solanacearum hrp genes ...

Southern Blot Chemiluminescent Detection System For Biotin

AuroraTM Southern Blotting kits are available in 2 sizes and sufficient for either 10 or 30 blots ( 10 cm x 10 cm). The reagents included in each kit are.

Isolation of a Gene Cluster from Xanthomonas campestris pv

pathogenicity are controlled by hrp (hypersensitive reaction. and pathogenicity; Niepold et al. exchange mutants were analyzed in Southern blot.

Cheluminate-HRP PicoDetect

Enhanced ChemiLuminescence Detection Kit for Western / Southern / Northern Blotting. Product-No. A3417. Description. The CheLuminate-HRP PicoDetect is a

FORENSIC BIOCHEMISTRY: SOUTHERN AND WESTERN BLOT

Southern blot is used for transferring. DNA the Northern blot for Southern blotting combines agarose gel ... in Western blots is HRP

Understanding secondary antibodies

HRP. Dot ELISA

Lumigen ECL Plus

blotted proteins. Lumigen ECL Plus utilizes a unique Western Blotting HRP Substrate ... This image from a Southern blot of a single copy gene of mouse.

[PDF] Cheluminate-HRP PicoDetect - ITW Reagents

The CheLuminate-HRP PicoDetect is a complete kit with ready-to-use reagents for chemiluminescent detection of immobilized proteins (Western blot) or

[PDF] KPL Detector™ HRP Chemiluminescent Blotting Kit - SeraCare

The KPL Detector HRP Chemiluminescent Blotting Kit was designed for the detection of plasmid and multiple-copy genomic Southern blots Using KPL LumiGLO

[PDF] SOUTHERN AND WESTERN BLOT TECHNIQUES DNA PROFILING

Southern blot is used for transferring DNA the Northern blot for RNA and the in Western blots is HRP a small stable enzyme with high specificity and

[PDF] Western- & Southern-Blotting Carl Roth

Transfer membrane for protein analyses Southern- and Northern-Blots Made of 100 nitrocellulose HRP- conjugate (rabbit) enables the marker to be

[PDF] Strip Northern And Southern Blots - Thermo Fisher Scientific

Following exposure of a Northern or Southern blot (either by classical isotopic 17097) test for stripping efficiency by reapplying streptavidin-HRP and

Short Technical Reports - Future Science

Schematic diagram of the sequential detection of CFTR genotypes with HRP and AP substrates Page 3 Short Technical Reports Southern Blot Analysis of CFTR

Preparation of horseradish peroxidase-labeled probes - PubMed

The direct labeling of nucleic acid probes with horseradish peroxidase (HRP) may including Southern blots Northern blots colony and plaque screening

[PDF] Western Blotting and ELISA Techniques - ResearchGate

The name Western blotting is a pun on the name Southern blotting a technique for DNA detection and the detection of RNA is termed northern blotting 2 1 Brief

[PDF] Strip Northern and Southern Blots

17095) test for stripping efficiency by re-applying streptavidin-HRP and add substrate as indicated in the protocol If no signal is detected then stripping

Methods Used to Detect Proteins and Nucleic Acids Bound to

HRP-Luminol system: The reaction between HRP and luminol can also be applied for detection of nucleic acids in Southern and Northern blotting

In the USA call: 800-8-PIERCE (800-874-3723) or 815-968-0747 • Fax: 815-968-7316 or 800-842-5007 • www.piercenet.com

Introduction

Following exposure of a Northern or Southern blot (either by classical isotopic methods or North2South

Chemiluminescent

Technology), researchers often wish to strip off the first probe and detecting reagents so that they may reprobe for the same

or different target. This Tech Tip describes several commonly used conditions for stripping probe from a Northern or

Southern blot while maintaining the nucleic acid target on the membrane (Table 1).

Be aware that any stripping procedure has the potential to denature sensitive nucleic acids, rendering them unrecognizable by

the probe. Should this occur, reprobing will not be possible, and a new blot will have to be prepared for each probing

experiment. Furthermore, any physical defects of a membrane will be exaggerated when stripped and reprobed.

Table 1. Possible stripping conditions for Northern and Southern blots.

Stripping Solution Strength DNA RNA Temperature & Time 0.5% SDS Mild Yes Yes 60°C for 60 minutes

5 mM Tris•HCl, pH 8.0, 2 mM EDTA,

0.1X Denhardt's Reagent Mild Yes Yes 65°C for 2 hours

50% Formamide, 2X SSPE Moderate Yes Yes 65°C for 1 hour

NaOH, 0.1% (w/v) SDS* Moderate Yes No 45°C for 30 minutes, then room temperature for 2 x 10 minutes Boiling 0.1% SDS Harsh Yes Yes Pour on 100°C solution and let cool to room temperature; repeat 2 times

0.1X SSPE or SSC/0.5% SDS Harsh Yes Yes Pour on 100°C solution and let cool to

room temperature; repeat 2 times *Blots must be neutralized in TE after stripping.

Tips for Stripping and Reprobing Blots

Wash blots with target side up

Handle blots only at corners

Keep blots wet between hybridization and stripping

Keep blots wet after stripping

Store blots in 0.1% SDS or 1X TE at 4°C

Stripped blots may be dried and stored at room temperature for long-term storage. However, any probe remaining on the

blot will become permanently bound once the blot is dried.

Test for Stripping Efficiency

For biotinylated probes detected using North2South Chemiluminescent Hybridization and Detection Kit (Product No.

17095), test for stripping efficiency by re-applying streptavidin-HRP and add substrate as indicated in the protocol. If no

signal is detected, then stripping was complete. If signal is detected, try a stronger stripping method.

For probes directly labeled with HRP and directly detected using North2South®

Direct HRP Labeling and Detection Kit

(Product No. 17195), add substrate as indicated in the protocol. If no signal is detected, then stripping was complete. If signal

is detected, try a stronger stripping method.

3747 N. Meridian Road

P.O. Box 117

Rockford, IL 61105

TR0028.0 Strip Northern and Southern Blots

In the USA call: 800-8-PIERCE (800-874-3723) or 815-968-0747 • Fax: 815-968-7316 or 800-842-5007 • www.piercenet.com

Buffer Formulations

50 X Denhardt's Reagent 20 X SSPE

5 gm Ficoll

5 gm Polyvinylpyrrolidone

5 gm BSA

Adjust volume to 500 ml 800 ml H

2 O

175.3 gm NaCl

27.6 g NaH

2 PO 4 •H 2 O pH to 7.4 with NaOH

Adjust volume to 1 L

20 X SSC TE, pH 7.4

800 ml H2O

175.3 gm NaCl

88.2 g sodium citrate

pH to 7.0 with NaOH Adjust volume to 1 L 10 mM Tris•HCl (pH 7.4)

1 mM EDTA (pH 8.0)

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Direct HRP Labeling and Detection Kit, for direct labeling of probes longer than 50 nucleotides and subsequent Northern or Southern blotting detection

89880 Chemiluminescent Nucleic Acid Detection Module

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[PDF] TECHNICAL RESOURCE - Strip Northern and Southern Blots