Analysis Note

ControlIncludes negative control mouse IgG antibody and control primers specific for human c-myc promoter.

Application

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).

Research CategoryEpigenetics & Nuclear Function

Use ChIPAb+ LEF1 Mouse Monoclonal Antibody validated in WB, ChIP for the detection of LEF1.

Research Sub CategoryChromatin BiologyTranscription Factors

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.

General description

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.

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

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

Quality

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.

Specificity

Recognizes LEF1 (lymphoid enhancer-binding factor 1).

Storage and Stability

1 year at -20°C from date of shipment

Target description

55-57 kDa