重组Anti-MMP14抗体[EP1264Y] (ab51074)
![Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074)](https://www.abcam.cn/ps/products/51/ab51074/Images/ab51074-383743-anti-mmp14-antibody-ep1264y-western-blot-a431-wt-a431-mmp14-knockout-hela-caco2.jpg "Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1264Y] to MMP14
- Suitable for: Flow Cyt, IHC-P, IP, WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
概述
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产品名称
Anti-MMP14抗体[EP1264Y]
参阅全部 MMP14 一抗 -
描述
兔单克隆抗体[EP1264Y] to MMP14 -
宿主
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P HumanIP HumanWB MouseRatHuman
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免疫原
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阳性对照
- IHC: Human breast carcinoma, endometrium carcinoma and kidney tissues. WB: Human fetal spleen, esophagus and lung cancer tissue lysates, mouse and rat spleen tissue lysates; A431 and Caco-2 whole cell lysate. Flow Cyt: A549 cells. IP: A431 cell lysate.
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常规说明
The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
性能
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形式
Liquid -
存放说明
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. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP1264Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab51074 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| 应用 | Species |
|---|---|
| Flow Cyt |
Human
|
| ICC/IF |
Human
|
| IHC-P |
Human
|
| IP |
Human
|
| WB |
Mouse
Rat
Human
|
| All applications |
Cow
|
| 应用 | Ab评论 | 说明 |
|---|---|---|
| Flow Cyt | (1) |
1/20.
|
| IHC-P | (4) |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
| IP |
1/20.
For unpurified use at 1/100. |
|
| WB | (7) |
1/5000. Predicted molecular weight: 66 kDa.Can be blocked with MMP14 peptide (ab185123).
For unpurified use at 1/2000. |
| ICC/IF | (6) |
Use at an assay dependent concentration.
|
| 说明 |
|---|
|
Flow Cyt
1/20. |
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IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
IP
1/20. For unpurified use at 1/100. |
|
WB
1/5000. Predicted molecular weight: 66 kDa.Can be blocked with MMP14 peptide (ab185123). For unpurified use at 1/2000. |
|
ICC/IF
Use at an assay dependent concentration. |
靶标
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功能
Seems to specifically activate progelatinase A. May thus trigger invasion by tumor cells by activating progelatinase A on the tumor cell surface. May be involved in actin cytoskeleton reorganization by cleaving PTK7. -
组织特异性
Expressed in stromal cells of colon, breast, and head and neck. Expressed in lung tumors. -
序列相似性
Belongs to the peptidase M10A family.
Contains 4 hemopexin-like domains. -
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
The precursor is cleaved by a furin endopeptidase. -
细胞定位
Membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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数据库链接
- Entrez Gene: 281915 Cow
- Entrez Gene: 4323 Human
- Entrez Gene: 17387 Mouse
- Entrez Gene: 81707 Rat
- Omim: 600754 Human
- SwissProt: Q9GLE4 Cow
- SwissProt: P50281 Human
- SwissProt: P53690 Mouse
see all -
别名
- Matrix metallopeptidase 14 (membrane inserted) antibody
- Matrix metalloproteinase 14 antibody
- Matrix metalloproteinase-14 antibody
see all
图片
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Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074)All lanes : Anti-MMP14 antibody [EP1264Y] (ab51074) at 1/2000 dilution
Lane 1 : A431 wild-type cell lysate
Lane 2 : A431 MMP14 knockout cell lysate
Lane 3 : Caco-2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 66 kDa
Observed band size: 54 kDa why is the actual band size different from the predicted?Lanes 1 - 3: Merged signal (red and green). Green - ab51074 observed at 54 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab51074 was shown to react with MMP14 in A431 wild-type cells in Western blot. Loss of signal was observed when MMP14 knockout sample was used. A431 wild-type and MMP14 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% Milk in TBS-T (0.1% Tween®) before incubation with ab51074 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074)](https://www.abcam.cn/ps/products/51/ab51074/Images/ab51074-273199-anti-mmp14-antibody-ep1264y-western-blot.jpg)
Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074)All lanes : Anti-MMP14 antibody [EP1264Y] (ab51074) at 1/2000 dilution (unpurified)
Lane 1 : Human fetal spleen tissue lysate
Lane 2 : Human lung cancer tissue lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 66 kDa
Observed band size: 60,63 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking/Diluting buffer and concentration 5% NFDM/TBST
63 kDa: pro-form; 60 kDa: active form.
MCF7 is a MMP14 negative or weakly expressed cell line (PMID: 25977338 and PMID: 19208838).
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![Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074)](https://www.abcam.cn/ps/products/51/ab51074/Images/ab51074-274269-anti-mmp14-antibody-ep1264y-western-blot.jpg)
Western blot - Anti-MMP14 antibody [EP1264Y] (ab51074)All lanes : Anti-MMP14 antibody [EP1264Y] (ab51074) at 1/5000 dilution (purified)
Lane 1 : Human fetal spleen lysate
Lane 2 : Human lung cancer lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat spleen lysate
Lane 5 : Human esophagus lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 66 kDa
Observed band size: 66 kDaBlocking and diluting buffer: 5% NFDM/TBST.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074)](https://www.abcam.cn/ps/products/51/ab51074/Images/ab51074-274270-anti-mmp14-antibody-ep1264y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium carcinoma tissue sections labeling MMP14 with purified ab51074 at 1/100 dilution (1.7 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counterstain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution.
PBS instead of the primary antibody was used as the negative control (inset).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074)](https://www.abcam.cn/ps/products/51/ab51074/Images/ab51074-62971-anti-mmp14-antibody-ep1264y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MMP14 antibody [EP1264Y] (ab51074)This image is courtesy of an anonymous Abreviewab51074 (unpurified) at 1/500 staining human kidney tissue sections by IHC-P.
The tissue was formaldehyde fixed and a heat mediated antigen retrieval step (in Tris/EDTA) was performed. The tissue was then blocked with serum and incubated with the primary antibody. A biotinylated donkey anti-rabbit IgG was used as the secondary.
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![Immunocytochemistry/ Immunofluorescence - Anti-MMP14 antibody [EP1264Y] (ab51074)](https://www.abcam.cn/ps/products/51/ab51074/Images/ab51074-314152-anti-mmp14-antibody-ep1264y-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-MMP14 antibody [EP1264Y] (ab51074)ab51074 staining MMP14 in HT-1080 (human fibrosarcoma epithelial cell) cells by ICC/IF (Immunocytochemistry/Immunofluorescence).
Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at 1/1000 dilution (0.2 μg/ml). An Alexa Fluor® 488 Goat anti-rabbit (ab150077) was used as the secondary antibody at 1/1000 dilution (2 μg/ml). Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594, ab195889) was used as the counterstain antibody at 1/200 dilution (2.5 μg/ml). DAPI was used as a nuclear counterstain. Confocal image showing cytoplasmic and weakly membranous staining in HT-1080 cell line.
Negative control (bottom panels): MCF7 PMID: 19208838.
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![Immunoprecipitation - Anti-MMP14 antibody [EP1264Y] (ab51074)](https://www.abcam.cn/ps/products/51/ab51074/Images/ab51074-274271-anti-mmp14-antibody-ep1264y-immunoprecipitation.jpg)
Immunoprecipitation - Anti-MMP14 antibody [EP1264Y] (ab51074)ab51074 (purified) at 1/20 dilution (2 µg) immunoprecipitating MMP14 in A431 (human epidermoid carcinoma) whole cell lysate.
Lane 1: A431 whole cell lysate 10ug
Lane 2: ab51074 + A431 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab51074 in A431 whole cell lysateFor western blotting, ab131366 VeriBlot for IP (HRP) was used for detection (1/1000).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
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![Flow Cytometry - Anti-MMP14 antibody [EP1264Y] (ab51074)](https://www.abcam.cn/ps/products/51/ab51074/Images/ab51074-314154-anti-mmp14-antibody-ep1264y-flow-cytometry.jpg)
Flow Cytometry - Anti-MMP14 antibody [EP1264Y] (ab51074)Flow cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell, Left) / HT-1080 (Human fibrosarcoma epithelial cell, Right) cells labeling MMP14 with ab51074 at 1/200 dilution (0.1 μg) (red). Goat anti-rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/2000 dilution. Rabbit monoclonal IgG (ab172730) / black was used as the isotype control. Cells incubated with secondary antibody only (blue) was used as the unlabeled control. Gated on viable cells.
Positive control (Right panel): HT-1080 cells.
Negative control (Left panel): MCF7 cells.
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![Anti-MMP14 antibody [EP1264Y] (ab51074)](https://www.abcam.cn/ps/products/51/ab51074/Images/ab51074-12-benefits-of-recombinant-antibodies.png)
Anti-MMP14 antibody [EP1264Y] (ab51074)
文献 (112)
ab51074 被引用在 112 文献中.
- Klepfish M et al. LOXL2 Inhibition Paves the Way for Macrophage-Mediated Collagen Degradation in Liver Fibrosis. Front Immunol 11:480 (2020). PubMed: 32296422
- Thuault S et al. A proximity-labeling proteomic approach to investigate invadopodia molecular landscape in breast cancer cells. Sci Rep 10:6787 (2020). PubMed: 32321993
- Gissi C et al. Extracellular vesicles from rat-bone-marrow mesenchymal stromal/stem cells improve tendon repair in rat Achilles tendon injury model in dose-dependent manner: A pilot study. PLoS One 15:e0229914 (2020). PubMed: 32163452
- Yao J et al. Single-cell transcriptomic analysis in a mouse model deciphers cell transition states in the multistep development of esophageal cancer. Nat Commun 11:3715 (2020). PubMed: 32709844
- Kasurinen A et al. High tissue MMP14 expression predicts worse survival in gastric cancer, particularly with a low PROX1. Cancer Med 8:6995-7005 (2019). PubMed: 31560170
