MMP-2 Recombinant Rabbit Monoclonal Antibody [PSH11-26]
cat.: HA723308
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: PSH11-26
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 74 kDa
Isotype: IgG
Immunogen: Recombinant protein within human MMP-2 aa 1-660.
Positive control: U-87 MG cell lysate, NIH/3T3 cell lysate, L6 cell lysate, Mouse skin tissue lysate, Rat skin tissue lysate, U-87 MG, NIH/3T3, L6, human colon tissue, human placenta tissue, human skin tissue, mouse colon tissue, mouse skin tissue, rat colon tissue, rat placenta tissue, rat skin tissue.
Subcellular location: Secreted, extracellular space, extracellular matrixm Membrane, Nucleus; Cytoplasm, Mitochondrion.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:2,000
1:100
1:1,000
1:1,000
1-2μg/sample
Uniprot #: SwissProt: P08253 Human | P33434 Mouse | P33436 Rat
Alternative names: 72 kDa gelatinase 72kD type IV collagenase CLG 4 CLG 4A CLG4 CLG4A Collagenase Type 4 alpha Collagenase type IV A Gelatinase A Gelatinase alpha Gelatinase neutrophil Matrix metallopeptidase 2 gelatinase A 72kDa gelatinase 72kDa type IV collagenase Matrix metalloproteinase 2 (gelatinase A, 72kDa gelatinase, 72kDa type IV collagenase) Matrix Metalloproteinase 2 Matrix metalloproteinase II Matrix metalloproteinase-2 MMP 2 MMP II MMP-2 MMP2 MMP2_HUMAN MONA Neutrophil gelatinase PEX TBE 1 TBE-1
Images
HA723308_1.jpg Fig1: Western blot analysis of MMP-2 on different lysates with Rabbit anti-MMP-2 antibody (HA723308) at 1/2,000 dilution.

Lane 1: U-87 MG cell lysate (15 µg/Lane)
Lane 2: HCT 116 cell lysate (low expression) (15 µg/Lane)
Lane 3: NIH/3T3 cell lysate (15 µg/Lane)
Lane 4: L6 cell lysate (15 µg/Lane)
Lane 5: Mouse skin tissue lysate (20 µg/Lane)
Lane 6: Rat skin tissue lysate (20 µg/Lane)

Predicted band size: 74 kDa
Observed band size: 70 kDa

Exposure time: 30 seconds; ECL: K1802;

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 (HA723308) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA723308_2.jpg Fig2: Immunocytochemistry analysis of U-87 MG (positive) and HCT 116 (low expression) labeling MMP-2 with Rabbit anti-MMP-2 antibody (HA723308) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MMP-2 antibody (HA723308) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA723308_3.jpg Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling MMP-2 with Rabbit anti-MMP-2 antibody (HA723308) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MMP-2 antibody (HA723308) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA723308_4.jpg Fig4: Immunocytochemistry analysis of L6 cells labeling MMP-2 with Rabbit anti-MMP-2 antibody (HA723308) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MMP-2 antibody (HA723308) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA723308_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-MMP-2 antibody (HA723308) 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 (HA723308) 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.
HA723308_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-MMP-2 antibody (HA723308) 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 (HA723308) 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.
HA723308_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-MMP-2 antibody (HA723308) 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 (HA723308) 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.
HA723308_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-MMP-2 antibody (HA723308) 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 (HA723308) 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.
HA723308_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse skin tissue with Rabbit anti-MMP-2 antibody (HA723308) 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 (HA723308) 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.
HA723308_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-MMP-2 antibody (HA723308) 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 (HA723308) 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.
HA723308_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded rat placenta tissue with Rabbit anti-MMP-2 antibody (HA723308) 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 (HA723308) 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.
HA723308_12.jpg Fig12: Immunohistochemical analysis of paraffin-embedded rat skin tissue with Rabbit anti-MMP-2 antibody (HA723308) 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 (HA723308) 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.
HA723308_13.jpg Fig13: Flow cytometric analysis of HCT 116 (left, low expression) and U-87 MG (right, positive) cells labeling MMP-2.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA723308, 1/1,000) (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 iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA723308_14.jpg Fig14: Flow cytometric analysis of NIH/3T3 cells labeling MMP-2.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA723308, 1/1,000) (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 iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA723308_15.jpg Fig15: MMP-2 was immunoprecipitated from 0.2 mg U-87 MG cell lysate with HA723308 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723308 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: U-87 MG cell lysate (input)
Lane 2: HA723308 IP in U-87 MG cell lysate
Lane 3: Rabbit IgG instead of HA723308 in U-87 MG cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 59 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.