Description
General description
55-57 kDa
Every lot of the ChIPAb+ line of antibodies is individually validated for chromatin precipitation, in order to guarantee successful ChIP assays every time. Each antibody includes a control primer set for performance confirmation. LEF1 antibody and the negative control antibody (mouse normal IgG) can be used to demonstrate that the LEF1 antibody is functionally validated in the precipitation of LEF1 associated chromatin.
The qPCR primers included flank the LEF1/TCF binding site in human c-myc promoter.
Immunogen
The antibody is made against amino acid residues 1-85 of human LEF1.
Application
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
Transcription Factors
Use ChIPAb+ LEF1 Mouse Monoclonal Antibody validated in WB, ChIP for the detection of LEF1.
Western Blot Analysis:
Jurkat cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-LEF1 (1 mg/mL). Proteins were visualized using a goat antimouse secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).
Biochem/physiol Actions
Recognizes LEF1 (lymphoid enhancer-binding factor 1).
Packaging
25 assays per kit. ~4μg per chromatin immunoprecipitation.
Physical form
Anti-LEF1 (mouse IgG1). 1 vial containing 100 mg of thiophilic and size exclusion chromatography purified mouse IgG1 in 100 mL of phosphate buffered saline with sodium azide. Store at-20°C. The antibody is made against amino acid residues 1-85 of human LEF1 and can recognize human and mouse LEF1. It does not cross-react with TCF-3 or TCF-4.
Normal Mouse IgG. One vial containing 125 μg of normal mouse IgG in 125 μL volume. Store at -20°C.
ChIP primers c-myc. One vial containing 75 mL of 5 μM of each control primer specific for human c-myc promoter. Store at -20°C.
FOR: CCC AAA AAA AGG CAC GGA A
REV: TAT TGG AAA TGC GGT CAT GC
Format: Purified
Preparation Note
1 year at -20°C from date of shipment
Analysis Note
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from 3x10⁶ HT29 cells was subjected to chromatin immunoprecipitation using 4 μg of either the negative control antibody, Mouse Normal IgG or mouse Anti-LEF1 and the Magna ChIP® G kit (Cat.# 17-611).
Successful enrichment of LEF1 associated DNA fragments was verified by qPCR using ChIP Primers c-myc flanking the human c-myc promoter that contains LEF1 binding sites (Please see figures).
Please refer to the EZ-Magna ChIP G (Cat.# 17-409) or EZ-ChIP (Cat.# 17-371) kit protocols for experimental details.
Control
Includes negative control mouse IgG antibody and control primers specific for human c-myc promoter.
Legal Information
MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
