MMP-9 Recombinant Rabbit Monoclonal Antibody [JA80-73]
cat.: ET1704-69
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JA80-73
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 78 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human MMP9 aa 71-120 / 707.
Positive control: THP-1 treated with 200nM TPA for 10 minutes whole cell lysate, rat lung tissue lysate, rat spleen tissue lysate, human tonsil tissue, A549-si-NT+TPA(80nM 24h) cell lysate, human spleen tissue, Hela, SHG-44, A431.
Subcellular location: Secreted.
Recommended Dilutions:
  WB
  IF-Tissue
  IHC-P
  FC
1:5,000
1:50-1:200
1:1,000
1:50-1:100
Uniprot #: SwissProt: P14780 Human | P41245 Mouse | P50282 Rat
Alternative names: 82 kDa matrix metalloproteinase-9 92 kDa gelatinase 92 kDa type IV collagenase CLG 4B CLG4B Collagenase Type 4 beta Collagenase type IV 92 KD EC 3.4.24.35 Gelatinase 92 KD Gelatinase B Gelatinase beta GelatinaseB GELB Macrophage gelatinase MANDP2 Matrix metallopeptidase 9 (gelatinase B, 92kDa gelatinase, 92kDa type IV collagenase) Matrix Metalloproteinase 9 MMP 9 MMP-9 MMP9 MMP9_HUMAN Type V collagenase
Images
ET1704-69_1.jpg Fig1: Western blot analysis of MMP-9 on different lysates with Rabbit anti-MMP-9 antibody (ET1704-69) at 1/5,000 dilution.

Lane 1: THP-1 whole cell lysate (20 µg/Lane)
Lane 2: THP-1 treated with 200nM TPA for 10 minutes whole cell lysate (20 µg/Lane)
Lane 3: Rat lung tissue lysate (20 µg/Lane)
Lane 4: Rat spleen tissue lysate (20 µg/Lane)

Predicted band size: 78 kDa
Observed band size: 92 kDa

Exposure time: 2 minutes;
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 (ET1704-69) at 1/5,000 dilution was used in 5% NFDM/TBST 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.
ET1704-69_2.jpg Fig2: Western blot analysis of MMP9 on different lysates with Rabbit anti-MMP9 antibody (ET1704-69) at 1/500 dilution.

Lane 1: A549-si NT+TPA(80nM 24h) cell lysate, 15 µg/Lane
Lane 2: A549-si MMP9#1+TPA(80nM 24h) cell lysate, 15 µg/Lane
Lane 3: A549-si MMP9#2+TPA(80nM 24h) cell lysate, 15 µg/Lane

Predicted band size: 78 kDa
Observed band size: 78-92 kDa

Exposure time: 1 minute 40 seconds;
4-20% SDS-PAGE gel.

ET1704-69 was shown to specifically react with MMP9 in Hela-si NT+TPA(80nM 24h) cells. No band was observed when Hela-si MMP9+TPA(80nM 24h) sample was tested. Hela-si NT+TPA(80nM 24h) and Hela-si MMP9+TPA(80nM 24h) 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 (ET1704-69, 1/500) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA 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.
ET1704-69_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-MMP-9 antibody (ET1704-69) 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 (ET1704-69) 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.
ET1704-69_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-MMP-9 antibody (ET1704-69) 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 (ET1704-69) 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.
ET1704-69_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-MMP-9 antibody (ET1704-69) 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 (ET1704-69) 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.
ET1704-69_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-MMP-9 antibody (ET1704-69) 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 (ET1704-69) 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.
ET1704-69_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-MMP-9 antibody (ET1704-69) 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 (ET1704-69) 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.
ET1704-69_8.jpg Fig8: Flow cytometric analysis of MMP-9 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1704-69, 1/50) (red). 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; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.