Recombinant Anti-MMP9 antibody [EPR22140-154] (ab228402)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22140-154] to MMP9
- Suitable for: IP, IHC-P, IHC-Fr, WB
- Reacts with: Mouse, Rat
Related conjugates and formulations
Overview
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Product name
Anti-MMP9 antibody [EPR22140-154]
See all MMP9 primary antibodies -
Description
Rabbit monoclonal [EPR22140-154] to MMP9 -
Host species
Rabbit -
Tested applications
Suitable for: IP, IHC-P, IHC-Fr, WBmore details -
Species reactivity
Reacts with: Mouse, Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse lung lysate; Rat lung, kidney, lymph node and spleen lysates. IHC-P: Mouse spleen and lung tissues; Rat spleen tissue. IHC-Fr: Mouse lung tissue. IP: Mouse lung lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22140-154 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab228402 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IP |
1/30.
|
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IHC-P | (1) |
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
IHC-Fr | (1) |
1/100.
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WB | (2) |
1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 81 kDa).
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Notes |
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IP
1/30. |
IHC-P
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IHC-Fr
1/100. |
WB
1/1000. Detects a band of approximately 92 kDa (predicted molecular weight: 81 kDa). |
Target
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Function
May play an essential role in local proteolysis of the extracellular matrix and in leukocyte migration. Could play a role in bone osteoclastic resorption. Cleaves KiSS1 at a Gly-
-Leu bond. Cleaves type IV and type V collagen into large C-terminal three quarter fragments and shorter N-terminal one quarter fragments. Degrades fibronectin but not laminin or Pz-peptide. -
Tissue specificity
Produced by normal alveolar macrophages and granulocytes. -
Involvement in disease
Intervertebral disc disease
Metaphyseal anadysplasia 2 -
Sequence similarities
Belongs to the peptidase M10A family.
Contains 3 fibronectin type-II domains.
Contains 4 hemopexin repeats. -
Domain
The conserved cysteine present in the cysteine-switch motif binds the catalytic zinc ion, thus inhibiting the enzyme. The dissociation of the cysteine from the zinc ion upon the activation-peptide release activates the enzyme. -
Post-translational
modificationsProcessing 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. -
Cellular localization
Secreted, extracellular space, extracellular matrix. - Information by UniProt
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Database links
- Entrez Gene: 17395 Mouse
- Entrez Gene: 81687 Rat
- SwissProt: P41245 Mouse
- SwissProt: P50282 Rat
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Alternative names
- 82 kDa matrix metalloproteinase-9 antibody
- 92 kDa gelatinase antibody
- 92 kDa type IV collagenase antibody
see all
Images
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All lanes : Anti-MMP9 antibody [EPR22140-154] (ab228402) at 1/1000 dilution
Lane 1 : Rat lung tissue lysate
Lane 2 : Rat brain tissue lysate
Lane 3 : Rat spleen tissue lysate
Lane 4 : Rat kidney tissue lysate
Lane 5 : Rat lymph node tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 81 kDa
Observed band size: 84-92 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking and diluting buffer and concentration:5% NFDM/TBST.
ab181602 was used as loading control for GAPDH.
Although MMP9 has been studied in brain in some publications, ab228402 was unable to detect signal in normal brain tissue, this may because MMP9 expression level is low in normal brain and would be increased in abnormal conditions like injury (PMID: 31198417)
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All lanes : Anti-MMP9 antibody [EPR22140-154] (ab228402) at 1/1000 dilution
Lane 1 : Mouse lung lysate
Lane 2 : Rat lung lysate
Lane 3 : Rat spleen lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 81 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted?Exposure times : Lanes 1-2: 3 minutes; Lane 3: 26 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
MMP9 is a glycoprotein (PMID: 29329315). The 92 kDa band likely represents the zymogen prior to activating cleavage of the pro-peptide (PMID: 7688350; PMID: 12901881).
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling MMP9 with ab228402 at 1/5,000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on polymorphonuclear leukocytes of mouse spleen (PMID: 28775117). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat-mediated antigen retrieval using citrate buffer (pH 6.0).
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Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse lung tissue labeling MMP9 with ab228402 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution (green). Positive staining on sporadic cells (macrophage-like) in mouse lung tissue is observed.
The nuclear counterstain is DAPI (blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution.
Heat-mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).
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MMP9 was immunoprecipitated from 0.35 mg of mouse brain lysate with ab228402 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab228402 at 1/1,000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution.
Lane 1: Mouse lung lysate 10 µg (Input).
Lane 2: ab228402 IP in mouse lung lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab228402 in mouse lung lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 20 seconds.
The blot was developed using a high sensitivity ECL.
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Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling MMP9 with ab228402 at 1/5,000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on polymorphonuclear leukocytes of mouse lung (PMID: 28775117). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat-mediated antigen retrieval using citrate buffer (pH 6.0).
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Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling MMP9 with ab228402 at 1/5,000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on polymorphonuclear leukocytes of rat spleen (PMID: 28775117). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat-mediated antigen retrieval using citrate buffer (pH 6.0).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (45)
ab228402 has been referenced in 45 publications.
- Li C et al. miR-362-3p suppresses ovarian cancer by inhibiting LRP8. Transl Oncol 15:101284 (2022). PubMed: 34839107
- Kim BH et al. Restoration of Homeostasis in the Tracheal Mucosa After Thyroid Surgery in a Rat Model. In Vivo 36:161-169 (2022). PubMed: 34972711
- Kim Y et al. HL3501, a Novel Selective A3 Adenosine Receptor Antagonist, Lowers Intraocular Pressure (IOP) in Animal Glaucoma Models. Transl Vis Sci Technol 11:30 (2022). PubMed: 35191964
- Liu P et al. Exosomes derived from stem cells of human deciduous exfoliated teeth inhibit angiogenesis in vivo and in vitro via the transfer of miR-100-5p and miR-1246. Stem Cell Res Ther 13:89 (2022). PubMed: 35241153
- Li YY et al. SCM-198 Alleviates Endometriosis by Suppressing Estrogen-ERα mediated Differentiation and Function of CD4+CD25+ Regulatory T Cells. Int J Biol Sci 18:1961-1973 (2022). PubMed: 35342349