MMP-9 Recombinant Rabbit Monoclonal Antibody [JA80-73] - BSA and Azide free
Catalog# HA750420
MMP-9 Recombinant Rabbit Monoclonal Antibody [JA80-73] - BSA and Azide free
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WB
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IF-Tissue
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IHC-P
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
MMP-9 Recombinant Rabbit Monoclonal Antibody [JA80-73] - BSA and Azide free
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
Target 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
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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.
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
ExpandImages
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☑ Cell treatment (CT)
This data was developed using ET1704-69, the same antibody clone in a different buffer formulation.
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. -
☑ Knockdown (KD)
This data was developed using ET1704-69, the same antibody clone in a different buffer formulation.
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. -
This data was developed using ET1704-69, the same antibody clone in a different buffer formulation.
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. -
This data was developed using ET1704-69, the same antibody clone in a different buffer formulation.
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. -
This data was developed using ET1704-69, the same antibody clone in a different buffer formulation.
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. -
This data was developed using ET1704-69, the same antibody clone in a different buffer formulation.
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. -
This data was developed using ET1704-69, the same antibody clone in a different buffer formulation.
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. -
This data was developed using ET1704-69, the same antibody clone in a different buffer formulation.
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. -
This data was developed using ET1704-69, the same antibody clone in a different buffer formulation.
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).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"