ETS Like-1 protein Elk-1 is a protein that in humans is encoded by the ELK1. Elk-1 functions as a transcription activator. It is classified as a ternary complex factor (TCF), a subclass of the ETS family, which is characterized by a common protein domain that regulates DNA binding to target sequences. Elk1 plays important roles in various contexts, including long-term memory formation, drug addiction, Alzheimer's disease, Down syndrome, breast cancer, and depression. Given its role as a transcription factor, Elk1 is expressed in the nuclei of non-neuronal cells. The protein is present in the cytoplasm as well as in the nucleus of mature neurons. In post-mitotic neurons, a variant of Elk1, sElk1, is expressed solely in the nucleus because it lacks the NES site present in the full-length protein. Moreover, while Elk1 is broadly expressed, actual levels vary among tissues. The rat brain, for example, is extremely rich in Elk1, but the protein is exclusively expressed in neurons.
Background References
1. Wei S et al. Elk1 enhances inflammatory cell infiltration and exacerbates acute lung injury/acute respiratory distress syndrome by suppressing Fcgr2b transcription. Mol Med. 2024 Apr
2. Wang J et al. ELK1 suppresses SYTL1 expression by recruiting HDAC2 in bladder cancer progression. Hum Cell. 2022 Nov
Western blot analysis of Phospho-ELK1 (S383) on different lysates with Rabbit anti-Phospho-ELK1 (S383) antibody (HA723663) at 1/5,000 dilution.
Lane 1: RAW264.7 cell lysate Lane 2: RAW264.7 treated with 10μg/mL LPS for 8 hours cell lysate Lane 3: RAW264.7 treated with 10μg/mL LPS for 8 hours cell lysate, then the membrane treated with λpp for 1 hour
Lysates/proteins at 20 µg/Lane.
Predicted band size: 45 kDa Observed band size: 55 kDa
Exposure time: 59 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723663) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human lung tissue untreated / treated with λpp with Rabbit anti-Phospho-ELK1 (S383) antibody (HA723663) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723663) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
☑ Cell treatment (CT)
Phospho-ELK1 (S383) was immunoprecipitated from 0.2 mg RAW264.7 treated with 0.1μg/mL LPS for 6 hours cell lysate with HA723663 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723663 at 1/25,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: RAW264.7 treated with 0.1μg/mL LPS for 6 hours cell lysate (input) Lane 2: HA723663 IP in RAW264.7 treated with 0.1μg/mL LPS for 6 hours cell lysate Lane 3: Rabbit IgG instead of HA723663 in RAW264.7 treated with 0.1μg/mL LPS for 6 hours cell lysate