MMP-12 Recombinant Rabbit Monoclonal Antibody [SR03-23]

Western blot analysis of MMP-12 on different lysates with Rabbit anti-MMP-12 antibody (ET1602-42) at 1/500 dilution.

Lane 1: Mouse lung tissue lysate
Lane 2: Rat lung tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 40 kDa

Exposure time: 1 minute;

8% 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 (ET1602-42) 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 1:200,000 dilution was used for 1 hour at room temperature.

☑ Knockdown (KD)

Western blot analysis of MMP12 on different lysates with Rabbit anti-MMP12 antibody (ET1602-42) at 1/1,000 dilution.

Lane 1: Hela-si NT cell lysate
Lane 2: Hela-si MMP12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 40 kDa

Exposure time: 6 minutes;

4-20% SDS-PAGE gel.

ET1602-42 was shown to specifically react with MMP12 in Hela-si NT cells. Weakened band was observed when Hela-si MMP12 sample was tested. Hela-si NT and Hela-si MMP12 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 (ET1602-42, 1/1,000) and Loading control antibody (Rabbit anti-HSP90, ET1605-56, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Western blot analysis of MMP-12 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 (ET1602-42, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: A549 cell lysate
Lane 2: Hela cell lysate
Lane 3: MCF-7 cell lysate
Lane 4: THP-1 cell lysate

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-MMP-12 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 primary antibody (ET1602-42, 1/50) for 30 minutes 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 mouse lung tissue using anti-MMP-12 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 primary antibody (ET1602-42, 1/50) for 30 minutes 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 MMP-12 was done on A549 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1602-42, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).