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Product Info Summary
| SKU: | A00605-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-LEF1 Antibody Picoband®
SKU/Catalog Number
A00605-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-LEF1 Antibody Picoband® catalog # A00605-2. Tested in ELISA, Flow Cytometry, IF, ICC, IHC, WB applications. This antibody reacts with Human, Mouse. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-LEF1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00605-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human LEF1 recombinant protein (Position: D14-H274).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00605-2 is reactive to LEF1 in Human, Mouse, Rat
Observed Molecular Weight
58 kDa
Calculated molecular weight
44.2 kDa
Background of LEF1
Lymphoid enhancer-binding factor 1 (LEF1) is a protein that in humans is encoded by the LEF1 gene. This gene encodes a transcription factor belonging to a family of proteins that share homology with the high mobility group protein-1. The protein encoded by this gene can bind to a functionally important site in the T-cell receptor-alpha enhancer, thereby conferring maximal enhancer activity. This transcription factor is involved in the Wnt signaling pathway, and it may function in hair cell differentiation and follicle morphogenesis. Mutations in this gene have been found in somatic sebaceous tumors. This gene has also been linked to other cancers, including androgen-independent prostate cancer. Alternative splicing results in multiple transcript variants.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00605-2 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human Jurkat whole cell, mouse thymus tissue, rat C6 whole cell
IHC: human tonsil tissue
ICC/IF: Caco-2 cell
FCM: HL-60 cell
Validation Images & Assay Conditions
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Western blot analysis of LEF1 using anti-LEF1 antibody (A00605-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: mouse thymus tissue lysates,
Lane 3: rat C6 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-LEF1 antigen affinity purified polyclonal antibody (Catalog # A00605-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for LEF1 at approximately 58 kDa. The expected band size for LEF1 is at 58 kDa.
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IHC analysis of LEF1 using anti-LEF1 antibody (A00605-2).
LEF1 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-LEF1 Antibody (A00605-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
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IF analysis of LEF1 using anti-LEF1 antibody (A00605-2).
LEF1 was detected in an immunocytochemical section of Caco-2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-LEF1 Antibody (A00605-2) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of HL-60 cells using anti-LEF1 antibody (A00605-2).
Overlay histogram showing HL-60 cells stained with A00605-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-LEF1 Antibody (A00605-2, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-LEF1 Antibody Picoband® (A00605-2)
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