Detection of Human LEF1 by Western Blot.
Western blot shows lysates of Jurkat human acute T cell leukemia cell line. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Human LEF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7647) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for LEF1 at approximately 35-55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
LEF1 in Neuro‑2A Mouse Cell Line.
LEF1 was detected in immersion fixed Neuro‑2A mouse neuroblastoma cell line using Goat Anti-Human LEF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7647) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
LEF1 (Lymphoid Enhancer-binding Factor 1) is a functionally diverse member of the TCF/LEF family of transcription factors. The name LEF1 was originally applied to the mouse factor, while its human counterpart was named TCF-1 alpha. Since there is a related molecule named TCF-1, to avoid confusion with TCF-1 alpha, both species factors are now called LEF1. Although its predicted MW is 44 kDa, it runs anomalously at 35-55 kDa in SDS-PAGE. LEF1 has restricted expression in adult, being limited to T cells, thymocytes, pre-B cells and NK cells. LEF1 demonstrates sequence-specific (CCTTTG[T/A][T/A]) DNA binding and directs beta -catenin to Wnt‑responsive genes. Either transcriptional activation or repression may occur on LEF1 target genes, depending upon 1) the cofactors recruited to the LEF1: beta ‑catenin complex, and 2) the phosphorylation state of LEF1. Human LEF1 is 399 amino acids (aa) in length. It contains one DNA-binding HMG-box (aa 298‑369), three potential phosphorylation sites, and an N-terminal SUMOylation site at Lys27. There are at least five isoform variants. One utilizes an alternative start site at Met116, a second contains a 47 aa substitution for aa 283‑299, and a third combines the previous two variations. A fourth variant shows a deletion of aa 214‑241, while a fifth variant combines the afore mentioned deletion of aa 214-241 with a 25 aa substitution for aa 390‑399. Over aa 120‑280, human LEF1 shares 97% aa sequence identity with mouse LEF1.
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