Biotin-labeled EMSA probe¯1„4‚‚β-Catenin-TCF

Aladdin's Biotin-labeled EMSA Probe - ²-Catenin/TCF is a ²-Catenin/TCF consensus oligonucleotide designed for EMSA . This double-stranded oligonucleotide contains the well-recognized binding sequence for ²-Catenin/TCF transcription factors, and can be used as a probe for EMSA. The nucleotide sequence of ²-Catenin/TCF consensus oligo is as followsPrecautions:Avoid heating above 40°C, as double-stranded DNA probe will be denatured at high temperatures into single strands that cannot be used in EMSA.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use:1.EMSA binding reactiona.Set up the following reactions by adding the reagents in sequence. Negative Control Reactions: Nuclease-Free Water7-7.5lEMSA/Gel-Shift Binding Buffer (5X)2lNuclear Protein or Purified Transcription Factor0lBiotin-labeled Probe0.5-1lTotal Volume10lSample Reaction:Nuclease-Free Water5-5.5lEMSA/Gel-Shift Binding Buffer (5X)2lNuclear Protein or Purified Transcription Factor2lBiotin-labeled Probe0.5-1lTotal Volume 10lCold Probe Competition Reaction:Nuclease-Free Water4-4.5lEMSA/Gel-Shift Binding Buffer (5X)2lNuclear Protein or Purified Transcription Factor2lUnlabeled Probe1lBiotin-labeled Probe0.5-1lTotal Volume 10lCold Mutant Probe Competition Reaction: Nuclease-Free Water4-4.5lEMSA/Gel-Shift Binding Buffer (5X)2lNuclear Protein or Purified Transcription Factor2lUnlabeled Probe1lBiotin-labeled Probe0.5-1lTotal Volume10lSuper-shift Reaction:Nuclease-Free Water4-4.5lEMSA/Gel-Shift Binding Buffer (5X)2lNuclear Protein or Purified Transcription Factor2lTarget Protein Specific Antibody1lBiotin-labeled Probe0.5-1lTotal Volume10lNote 1: The recommended dosage of biotin-labeled EMSA probe is 0.5l per reaction. If the EMSA bands of target proteins are weak, the dosage of biotin-labeled EMSA probe can be increased to 0.75 l or 1 l as appropriate. Note 2: For cold probe competition, the recommended amount of unlabeled probe is 50-100 times that of the biotin-labeled probe, but can be adjusted as required.b.Before adding the labeled probe, the reaction mix should be mixed well and incubated at room temperature (20-25°C) for 10 minutes to eliminate possible non-specific binding between probes and proteins. Otherwise allow cold probes react preferentially. Then add the labeled probe, mix well and incubate at room temperature (20-25°C) for 20 minutes.c.Add 1l of EMSA/Gel-Shift Loading Buffer (colorless, 10X) into each reaction and load onto gel immediately after mixing. Note: Sometimes bromophenol blue can affect protein-DNA interactions. It is recommended to use the colorless EMSA/Gel-Shift Loading Buffer, or that including a very small amount of blue loading buffer to make observation easier.2.For more detailed EMSA procedures with the biotin-labeled probes, please refer to the manual of 's Chemiluminescent EMSA Kit .

Specification: 0.2M

UPC:

41116126

Condition:

New

Weight:

0.07 Ounces

HazmatClass:

No

Quantity:

1

Unit of Measurement:

EA

WeightUOM:

LB

MPN:

B752138-200μl

Product Size:

200μl