ERK2 Mouse Monoclonal Antibody [5-D2]

Western blot analysis of ERK2 on different lysates with Mouse anti-ERK2 antibody (M1407-3) at 1/1,000 dilution.

Lane 1: Human brain tissue lysate
Lane 2: Mouse brain tissue lysate
Lane 3: Rat brain tissue lysate

Lysates/proteins at 40 µg/Lane.

Predicted band size: 41 kDa
Observed band size: 41 kDa

Exposure time: 1 minute 2 seconds;

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 (M1407-3) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Alpaca Anti-Mouse IgG - HRP for IP Nano-Secondary Antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

ICC staining ERK2 in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with ERK2 monoclonal antibody at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution.

ICC staining ERK2 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with ERK2 monoclonal antibody at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution.

ICC staining ERK2 in NIH-3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with ERK2 monoclonal antibody at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluorc™ 488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution.

Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-ERK2 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the antibody (M1407-3) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue using anti-ERK2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the antibody (M1407-3) at 1/100 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

Flow cytometric analysis of ERK2 was done on Hela cells. The cells were fixed, permeabilized and stained with ERK2 antibody at 1/100 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-mouse IgG Secondary antibody at 1/500 dilution.