TEAD1 Recombinant Rabbit Monoclonal Antibody [JE52-71]

☑ Knockout (KO)

All lanes: Western blot analysis of TEF1 with anti-TEF1 antibody [JE52-71] (ET7111-04) at 1:500 dilution.

Lane 1: Wild-type ct26.wt whole cell lysate.
Lane 2: TEF1 knockout ct26.wt whole cell lysate.

ET7111-04 was shown to specifically react with TEF1 in wild-type ct26.wt cells. No band was observed when TEF1 knockout samples were tested. Wild-type and TEF1 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-TEF1 antibody (ET7111-04, 1/500) and Anti-HSP90 antibody (ET1605-56, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Cell lysate was provided by Ubigene Biosciences (Ubigene Biosciences Co., Ltd., Guangzhou, China).

Western blot analysis of TEAD1 on different lysates with Rabbit anti-TEAD1 antibody (ET7111-04) at 1/1,000 dilution.

Lane 1: A431 cell lysate
Lane 2: A549 cell lysate
Lane 3: HeLa cell lysate
Lane 4: Mouse embryo tissue lysate
Lane 5: Mouse skeletal muscle tissue lysate
Lane 6: Rat embryo tissue lysate
Lane 7: Rat skeletal muscle tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 48 kDa
Observed band size: 48/50 kDa

Exposure time: 2 minutes 7 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 (ET7111-04) at 1/1,000 dilution was used in 5% NFDM/TBST 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 colon cancer tissue with Rabbit anti-TEAD1 antibody (ET7111-04) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET7111-04) at 1/200 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.

Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-TEAD1 antibody (ET7111-04) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (ET7111-04) at 1/200 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.

Immunocytochemistry analysis of A549 cells labeling TEAD1 with Rabbit anti-TEAD1 antibody (ET7111-04) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-TEAD1 antibody (ET7111-04) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Immunocytochemistry analysis of MG-63 cells labeling TEAD1 with Rabbit anti-TEAD1 antibody (ET7111-04) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-TEAD1 antibody (ET7111-04) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Immunocytochemistry analysis of SiHa cells labeling TEAD1 with Rabbit anti-TEAD1 antibody (ET7111-04) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-TEAD1 antibody (ET7111-04) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃.Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

TEAD1 was immunoprecipitated from 0.2 mg A549 cell lysate with ET7111-04 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET7111-04 at 1/1,000 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: A549 cell lysate (input)
Lane 2: ET7111-04 IP in A549 cell lysate
Lane 3: Rabbit IgG instead of ET7111-04 in A549 cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 3 minutes; ECL: K1801