Biotin-labeled EMSA probe－β-Catenin-TCF

Aladdin's Biotin-labeled EMSA Probe - β-Catenin/TCF is a β-Catenin/TCF consensus oligonucleotide designed for EMSA . This double-stranded oligonucleotide contains the well-recognized binding sequence for β-Catenin/TCF transcription factors, and can be used as a probe for EMSA. The nucleotide sequence of β-Catenin/TCF consensus oligo is as followsPrecautions：Avoid heating above 40℃, 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 Water7-7.5μlEMSA/Gel-Shift Binding Buffer (5X)2μlNuclear Protein or Purified Transcription Factor0μlBiotin-labeled Probe0.5-1μlTotal Volume10μlSample Reaction:Nuclease-Free Water5-5.5μlEMSA/Gel-Shift Binding Buffer (5X)2μlNuclear Protein or Purified Transcription Factor2μlBiotin-labeled Probe0.5-1μlTotal Volume 10μlCold Probe Competition Reaction:Nuclease-Free Water4-4.5μlEMSA/Gel-Shift Binding Buffer (5X)2μlNuclear Protein or Purified Transcription Factor2μlUnlabeled Probe1μlBiotin-labeled Probe0.5-1μlTotal Volume 10μlCold Mutant Probe Competition Reaction: Nuclease-Free Water4-4.5μlEMSA/Gel-Shift Binding Buffer (5X)2μlNuclear Protein or Purified Transcription Factor2μlUnlabeled Probe1μlBiotin-labeled Probe0.5-1μlTotal Volume10μlSuper-shift Reaction:Nuclease-Free Water4-4.5μlEMSA/Gel-Shift Binding Buffer (5X)2μlNuclear Protein or Purified Transcription Factor2μlTarget Protein Specific Antibody1μlBiotin-labeled Probe0.5-1μlTotal Volume10μlNote 1: The recommended dosage of biotin-labeled EMSA probe is 0.5µl per reaction. If the EMSA bands of target proteins are weak, the dosage of biotin-labeled EMSA probe can be increased to 0.75 µl or 1 µl 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℃) 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℃) for 20 minutes.c.Add 1μl 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: 0.2μM