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MMP-9 Recombinant Rabbit Monoclonal Antibody [JA80-73]

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Specification

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Catalog# ET1704-69

MMP-9 Recombinant Rabbit Monoclonal Antibody [JA80-73]

Application

  • WB

  • IF-Tissue

  • IHC-P

  • FC

  • mIHC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

MMP-9 Recombinant Rabbit Monoclonal Antibody [JA80-73]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human MMP9 aa 71-120 / 707.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Tissue, IHC-P, FC, mIHC

Molecular Weight

Predicted band size: 78 kDa

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.

Conjugation

unconjugated

Clone Number

JA80-73

RRID

AB_3070515

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000-1:10,000

  • IF-Tissue

  • 1:200-1:500

  • IHC-P

  • 1:1,000

  • FC

  • 1:500-1:1,000

  • mIHC

  • 1:1,000

Target

Function

The matrix metalloproteinases (MMPs) are a family of peptidase pathway responsible for the degradation of extracellular matrix components, including collagen, gelatin, fibronectin, laminin and proteoglycan. Transcription of MMP genes is is differentially activated by phorbol ester, lipopolysaccharide (LPS) or staphylococcal enterotoxin MMP-9 (also designated 92 kDa type IV collagenase or gelatinase B) has been shown to degrade bone collagens in concert with MMP-1 (also specified interstitial collagenase, fibroblast collagenase or Collagenase-1), and cysteine proteases and may play a role in bone osteoclastic resorption. MMP-1 is downregulated by p53, and abnormality of p53 expression can contribute to joint degradation in rheumatoid arthritis by regulating MMP-1 expression.

Background References

1. Xu L et al. Umbilical cord-derived mesenchymal stem cells on scaffolds facilitate collagen degradation via upregulation of MMP-9 in rat uterine scars. Stem Cell Res Ther 8:84 (2017).

2. Gong L et al. Transthyretin regulates the migration and invasion of JEG-3 cells. Oncol Lett 13:1242-1246 (2017).

Sequence Similarity

Belongs to the peptidase M10A family.

Tissue Specificity

Detected in neutrophils (at protein level). Produced by normal alveolar macrophages and granulocytes.

Post-translational Modification

Processing of the precursor yields different active forms of 64, 67 and 82 kDa. Sequentially processing by MMP3 yields the 82 kDa matrix metalloproteinase-9.; N- and O-glycosylated.

Subcellular Location

Secreted.

UNIPROT

SwissProt: P14780 Human

SwissProt: P41245 Mouse

SwissProt: P50282 Rat

Synonyms

82 kDa matrix metalloproteinase-9 antibody

92 kDa gelatinase antibody

92 kDa type IV collagenase antibody

CLG 4B antibody

CLG4B antibody

Collagenase Type 4 beta antibody

Collagenase type IV 92 KD antibody

EC 3.4.24.35 antibody

Gelatinase 92 KD antibody

Gelatinase B antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of MMP-9 on different lysates with Rabbit anti-MMP-9 antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) at 1/5,000 dilution.<br /><br />Lane 1: THP-1 whole cell lysate (20 µg/Lane)<br />Lane 2: THP-1 treated with 200nM TPA for 10 minutes whole cell lysate (20 µg/Lane)<br />Lane 3: Rat lung tissue lysate (20 µg/Lane)<br />Lane 4: Rat spleen tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 78 kDa<br />Observed band size: 92 kDa<br /><br />Exposure time: 2 minutes;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of MMP9 on different lysates with Rabbit anti-MMP9 antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) at 1/5,000 dilution.<br /><br />Lane 1: A549-si NT+TPA(80nM 24h) cell lysate, 15 µg/Lane<br />Lane 2: A549-si MMP9#1+TPA(80nM 24h) cell lysate, 15 µg/Lane<br />Lane 3: A549-si MMP9#2+TPA(80nM 24h) cell lysate, 15 µg/Lane<br /><br />Predicted band size: 78 kDa<br />Observed band size: 78-92 kDa<br /><br />Exposure time: 1 minute 40 seconds;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>, 1/5,000) and Loading control antibody (Rabbit anti-GAPDH, <a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/100,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of MMP9 on different lysates with Rabbit anti-MMP9 antibody (ET1704-69) at 1/5,000 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.

    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, 1/5,000) 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 1/100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-MMP-9 antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-MMP-9 antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-MMP-9 antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-MMP-9 antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-MMP-9 antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>) 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.

    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.

  • Application: Immunofluorescence (IF-tissue)<br /><br />Species: Human<br />Tissue: Spleen<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× PBS<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>, 1/500, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IF-tissue)

    Species: Human
    Tissue: Spleen
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× PBS
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: ET1704-69, 1/500, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.

  • Flow cytometric analysis of MMP-9 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1704-69" style="font-weight: bold;text-decoration: underline;">ET1704-69</a>, 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).

    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).

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Citation

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