重组Anti-MMP9抗体[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
概述
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产品名称
Anti-MMP9抗体[EP1254] - BSA and Azide free
参阅全部 MMP9 一抗 -
描述
兔单克隆抗体[EP1254] to MMP9 - BSA and Azide free -
宿主
Rabbit -
特异性
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. -
经测试应用
适用于: ICC/IF, IHC-P, WB, Flow Cyt (Intra)more details -
种属反应性
与反应: Rat, Human, Recombinant fragment -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: U937, HL60 and TPA treated HT1080 cell lysates, human and rat lung, spleen and lymph node 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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常规说明
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
解离常数(KD)
KD = 1.58 x 10 -10 M Learn more about KD -
存储溶液
pH: 7.20
Constituent: 100% PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP1254 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-MMP9 antibody [EP1254] (ab309755)
- Alexa Fluor® 647 Anti-MMP9 antibody [EP1254] (ab310122)
- Alexa Fluor® 594 Anti-MMP9 antibody [EP1254] (ab310533)
- Alexa Fluor® 555 Anti-MMP9 antibody [EP1254] (ab312063)
- Alexa Fluor® 568 Anti-MMP9 antibody [EP1254] (ab312540)
- Anti-MMP9 antibody [EP1254] (ab76003)
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab204850于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P | (1) |
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. |
|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
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说明 |
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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. |
靶标
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功能
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. -
组织特异性
Produced by normal alveolar macrophages and granulocytes. -
疾病相关
Intervertebral disc disease
Metaphyseal anadysplasia 2 -
序列相似性
Belongs to the peptidase M10A family.
Contains 3 fibronectin type-II domains.
Contains 4 hemopexin repeats. -
结构域
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. -
翻译后修饰
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. -
细胞定位
Secreted, extracellular space, extracellular matrix. - Information by UniProt
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数据库链接
- Entrez Gene: 4318 Human
- Entrez Gene: 81687 Rat
- Omim: 120361 Human
- SwissProt: P14780 Human
- SwissProt: P50282 Rat
- Unigene: 297413 Human
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别名
- 82 kDa matrix metalloproteinase-9 antibody
- 92 kDa gelatinase antibody
- 92 kDa type IV collagenase antibody
see all
图片
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All lanes : Anti-MMP9 antibody [EP1254] (ab76003) at 1/1000 dilution
Lane 1 : Human lung tissue lysate
Lane 2 : Human brain tissue lysate
Lane 3 : Human spleen tissue lysate
Lane 4 : Human lymph node tissue lysate
Lane 5 : Rat lung tissue lysate
Lane 6 : Rat brain tissue lysate
Lane 7 : Rat spleen tissue lysate
Lane 8 : Rat kidney tissue lysate
Lane 9 : 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: 78 kDa
Observed band size: 84-92 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76003).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds; 40 seconds.
ab181602 was used as loading control for GAPDH.
Although MMP9 has been studied in brain in some publications, ab76003 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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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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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) 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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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.
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (1)
ab204850 被引用在 1 文献中.
- McDowell SAC et al. Neutrophil oxidative stress mediates obesity-associated vascular dysfunction and metastatic transmigration. Nat Cancer 2:545-562 (2021). PubMed: 35122017