Desmin Mouse Monoclonal Antibody [3-F7]

Western blot analysis of Desmin on different lysates with Mouse anti-Desmin antibody (M1501-10) at 1/1,000 dilution.

Lane 1: Rat heart tissue lysate
Lane 2: Rat skeletal muscle tissue lysate

Lysates/proteins at 40 µg/Lane.

Predicted band size: 53 kDa
Observed band size: 53 kDa

Exposure time: 3 minutes; 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 (M1501-10) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Western blot analysis of Desmin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Human skeletal muscle tissue lysate, untreated
Lane 2: Human heart tissue lysate, untreated

☑ Knockout (KO)

All lanes: Western blot analysis of Desmin with anti-Desmin antibody (M1501-10) at 1:500 dilution.
Lane 1: Wild-type Hela whole cell lysate (10 µg).
Lane 2/3: Desmin knockout Hela whole cell lysate (10 µg).

M1501-10 was shown to specifically react with Desmin in wild-type Hela cells. NO bands were observed when Desmin knockout sample were tested. Wild-type and Desmin 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 antibody (M1501-10, 1:500) was used in 5% BSA at room temperature for 2 hours. Goat anti-Mouse IgG-HRP antibody (HA1006) at 1:50,000 dilution was used for 1 hour at room temperature.

ICC staining Desmin in D3 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 Desmin 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 Desmin in Hela 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 Desmin 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. The nuclear counter stain is DAPI (blue).

ICC staining Desmin 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 Desmin 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 cervix tissue using anti-Desmin 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 (M1501-10) 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 stomach cancer tissue using anti-Desmin 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 (M1501-10) 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 uterus tissue using anti-Desmin 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 (M1501-10) 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 Desmin was done on Hela cells. The cells were fixed, permeabilized and stained with Desmin 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 for 30 minutes.