Recombinant Anti-MMP9 antibody [EP1254] - BSA and Azide free (ab204850)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1254] to MMP9 - BSA and Azide free
- Suitable for: ICC/IF, IHC-P, WB, Flow Cyt (Intra)
- Reacts with: Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-MMP9 antibody [EP1254] - BSA and Azide free
See all MMP9 primary antibodies -
Description
Rabbit monoclonal [EP1254] to MMP9 - BSA and Azide free -
Host species
Rabbit -
Specificity
Based on our preliminary data, ab204850 detects no or weak band of interest in the untreated cell lines at the dilution of 1/200. Treatment increasing the expression of MMP-9 is recommended when using this antibody. -
Tested applications
Suitable for: ICC/IF, IHC-P, WB, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Rat, Human, Recombinant fragment -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: U937, HL60 and TPA treated HT1080 cell lysates and rat kidney tissue lysate. IHC-P: Human gastic adenocarcinoma and spleen tissue. ICC/IF: U-2 OS and domoic acid-treated U87-MG cells. Flow Cyt (intra): A431 cells.
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General notes
ab204850 is the carrier-free version of ab76003.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Dissociation constant (KD)
KD = 1.58 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Constituent: 49% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1254 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab204850 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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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 78 kDa.
Based on our preliminary data, ab204850 detects no or weak band of interest in the untreated cell lines at the dilution of 1/200. Treatment increasing the expression of MMP-9 is recommended when using this antibody. |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 78 kDa. Based on our preliminary data, ab204850 detects no or weak band of interest in the untreated cell lines at the dilution of 1/200. Treatment increasing the expression of MMP-9 is recommended when using this antibody. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
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: 4318 Human
- Entrez Gene: 81687 Rat
- Omim: 120361 Human
- SwissProt: P14780 Human
- SwissProt: P50282 Rat
- Unigene: 297413 Human
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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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Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [EP1254] - BSA and Azide free (ab204850)
Immunocytochemistry/Immunofluorescence analysis of U-2 OS (human osteosarcoma) cells labeling MMP9 with ab76003 at 1/500 (4.3 μg/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody. Cells were counterstained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200, 2.5 μg/mL) - Microtubule Marker (Alexa Fluor® 594). DAPI (blue) was used as a nuclear counterstain.
Confocal image showing cytoplasmic staining onU-2 OS cells, the expression increased after treatment with TPA (200 nM) for 24 hours (middle panel).
Secondary Only Control: PBS was used instead of the primary antibody as the negative control with both TPA treated and untreated U-2 OS cells.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76003).
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Immunocytochemistry/ Immunofluorescence - Anti-MMP9 antibody [EP1254] - BSA and Azide free (ab204850)
Unpurified ab76003 staining MMP9 in U87-MG cells treated with domoic acid (ab120338), by ICC/IF. Increase of MMP9 expression correlates with increased concentration of domoic acid, as described in literature.
The cells were incubated at 37°C for 6h in media containing different concentrations of ab120338 (domoic acid) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with unpurified ab76003 (1/200) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight® 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76003).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] - BSA and Azide free (ab204850)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric adenocarcinoma tissue labeling MMP9 with unpurified ab76003 at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76003).
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Overlay histogram showing permeabilized A431 (Human epidermoid carcinoma cell line) cells stained with unpurified ab76003 (pink line).
Negative control antibody (green line) was rabbit IgG.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76003).
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Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76003).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP9 antibody [EP1254] - BSA and Azide free (ab204850)
This IHC data was generated using the same anti-MMP9 antibody clone, EP1254, in a different buffer formulation (cat# ab76003).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling MMP9 with purified ab76003 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab204850 has not yet been referenced specifically in any publications.