TPMT Recombinant Rabbit Monoclonal Antibody [JE64-09]

Western blot analysis of TPMT on different lysates with Rabbit anti-TPMT antibody (HA721049) at 1/500 dilution.

Lane 1: K562 cell lysate
Lane 2: SK-Br-3 cell lysate
Lane 3: HepG2 cell lysate
Lane 4: A431 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 28 kDa
Observed band size: 28 kDa

Exposure time: 2 minutes;

12% 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 (HA721049) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Western blot analysis of TPMT on different lysates with Rabbit anti-TPMT antibody (HA721049) at 1/500 dilution.

Lane 1: Mouse brain tissue lysate
Lane 2: Rat stomach tissue lysate
Lane 3: Mouse kidney tissue lysate
Lane 4: Rat lung tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 28 kDa
Observed band size: 28 kDa

Exposure time: 2 minutes;

12% 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 (HA721049) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue with Rabbit anti-TPMT antibody (HA721049) at 1/400 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 (HA721049) at 1/400 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.

Flow cytometric analysis of K562 cells labeling TPMT.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721049, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a Alexa Fluor® 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-TPMT antibody (HA721049) 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 (HA721049) 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.

☑ Knockdown (KD)

Western blot analysis of TPMT on different lysates with Rabbit anti-TPMT antibody (HA721049) at 1/2,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-TPMT KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 28 kDa
Observed band size: 28 kDa

Exposure time: 120 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 (HA721049) at 1/2,000 dilution was used in 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.