Western blot analysis of MMP3 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 (bsm-54152R, 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: human liver tissue lysate Lane 2: rat liver tissue lysate

Blocking buffer: 5% NFDM/TBST
Primary ab dilution: 1:1000
Primary ab incubation condition: 2 hours at
room temperature
Secondary ab: Goat Anti-Rabbit IgG H&L
(HRP)
Lysate: 1: Raji, 2: U87-MG, 3: Mouse placenta,
4: Rat placenta
Protein loading quantity: 20 μg
Exposure time: 60 s
Predicted MW: 54 kDa
Observed MW: 54 kDa

Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-MMP3 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 (bsm-54152R, 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 human liver tissue using anti-MMP3 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 (bsm-54152R, 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 rat kidney tissue using anti-MMP3 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 (bsm-54152R, 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 human placenta tissue using anti-MMP3 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 (bsm-54152R, 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.

Tissue: Human liver
Section type: Formalin fixed & Paraffin -
embedded section
Retrieval method: High temperature and high
pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary ab dilution: 1:100
Primary ab incubation condition: 1 hour at
room temperature
Secondary ab: Anti-Rabbit and Mouse
Polymer HRP (Ready to use)
Counter stain: Hematoxylin (Blue)
Comment:
Color brown is the positive signal
for bsm-54152R

Cell line: HT-29
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% TritonX-100
Primary ab dilution: 1:50
Primary incubation condition: 4°C overnight
Secondary ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (Blue)
Comment: Color green is the positive signal for bsm-54152R

Cell line: NIH/3T3
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% TritonX-100
Primary ab dilution: 1:50
Primary incubation condition: 4°C overnight
Secondary ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (Blue)
Comment: Color green is the positive signal for bsm-54152R

Cell line: HT-29
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% TritonX-100
Primary ab dilution: 1:50
Primary incubation condition: 4°C overnight
Secondary ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (Blue)
Comment: Color green is the positive signal for
bsm-54152R

Cell line: NIH/3T3
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% TritonX-100
Primary ab dilution: 1:50
Primary incubation condition: 4°C overnight
Secondary ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (Blue)
Comment: Color green is the positive signal for
bsm-54152R

Cell line: HepG2
Fixation: 4% Paraformaldehyde
Permeabilization: 90% Methanol
Primary Ab dilution: 1:100
Secondary Ab: Goat Anti-Rabbit IgG
Unlabelled control: The cell without incubation with primary antibody and secondary antibody (Black line).
Isotype control: Rabbit monoclonal IgG (Blue line).
Comment: Line red is the positive signal for bsm-54152R

Cell line: HepG2
Fixation: 4% Paraformaldehyde
Permeabilization: 90% Methanol
Primary Ab dilution: 1:100
Secondary Ab: Goat Anti-Rabbit IgG
Unlabelled control: The cell without incubation
with primary antibody and secondary antibody
(Black line).
Isotype control: Rabbit monoclonal IgG (Blue
line).
Comment: Line red is the positive signal for

bsm-54152R