Anti-MMP14 抗体 [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: IHC-P, IP, Flow Cyt (Intra), WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-MMP14 antibody [EP1264Y]
MMP14 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1264Y] to MMP14 -
由来種
Rabbit -
アプリケーション
適用あり: IHC-P, IP, Flow Cyt (Intra), WB, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Cow -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab185123) -
ポジティブ・コントロール
- IHC-P: Human endometrium carcinoma and kidney tissues. WB: Human fetal spleen, esophagus and lung cancer tissue lysates, mouse and rat spleen tissue lysates; A431 wild-type cell lysate and Caco-2 whole cell lysate. Flow Cyt (intra): HT-1080 cells. IP: A431 cell lysate. ICC/IF: HT-1080 cells and wild-type A431 cells.
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP1264Y -
アイソタイプ
IgG -
研究分野
関連製品
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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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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab51074の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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IHC-P | (4) |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/20.
For unpurified use at 1/100. |
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Flow Cyt (Intra) |
1/200.
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WB | (8) |
1/5000. Predicted molecular weight: 66 kDa.Can be blocked with MMP14 peptide (ab185123).
For unpurified use at 1/2000. |
ICC/IF | (6) |
Use a concentration of 0.2 µg/ml.
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特記事項 |
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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. |
Flow Cyt (Intra)
1/200. |
WB
1/5000. Predicted molecular weight: 66 kDa.Can be blocked with MMP14 peptide (ab185123). For unpurified use at 1/2000. |
ICC/IF
Use a concentration of 0.2 µg/ml. |
ターゲット情報
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機能
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. -
組織特異性
Expressed in stromal cells of colon, breast, and head and neck. Expressed in lung tumors. -
配列類似性
Belongs to the peptidase M10A family.
Contains 4 hemopexin-like domains. -
ドメイン
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. -
翻訳後修飾
The precursor is cleaved by a furin endopeptidase. -
細胞内局在
Membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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参照データベース
- 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 -
別名
- Matrix metallopeptidase 14 (membrane inserted) antibody
- Matrix metalloproteinase 14 antibody
- Matrix metalloproteinase-14 antibody
see all
画像
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All lanes : Anti-MMP14 antibody [EP1264Y] (ab51074) at 1/2000 dilution
Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : MMP14 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : Caco-2 (Human colorectal adenocarcinoma cell line) whole 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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ab51074 staining MMP14 in wild-type A431 cells (top panel) and MMP14 knockout A431 cells (bottom panel) (ab261890). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab51074 at 0.2μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
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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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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Immunohistochemical analysis of paraffin-embedded Human endometrial cancer labeling MMP14 with ab51074 at 1/500 (3.26 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Postive staining on the human endometrial tumour stroma cells and weak staining on the tumour cells. The section was incubated with ab51074 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. -
Immunohistochemical analysis of paraffin-embedded Human endometrial cancer labeling MMP14 with ab51074 at 1/500 (3.26 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human endometrial cancer. The section was incubated with ab51074 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.
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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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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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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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Intracellular 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 Fluorr® 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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ab51074 (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.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (175)
ab51074 は 175 報の論文で使用されています。
- Zhu L et al. A Zeb1/MtCK1 metabolic axis controls osteoclast activation and skeletal remodeling. EMBO J 42:e111148 (2023). PubMed: 36843552
- Kato T et al. Interplay of adherens junctions and matrix proteolysis determines the invasive pattern and growth of squamous cell carcinoma. Elife 12:N/A (2023). PubMed: 36892272
- Lin CP et al. Tributyrin Intake Attenuates Angiotensin II-Induced Abdominal Aortic Aneurysm in LDLR-/- Mice. Int J Mol Sci 24:N/A (2023). PubMed: 37175712
- Ku M & Yang J Intracellular lipophilic network transformation induced by protease-specific endocytosis of fluorescent Au nanoclusters. Nano Converg 10:26 (2023). PubMed: 37296273
- Zhang J et al. Genetically engineered nano-melittin vesicles for multimodal synergetic cancer therapy. Bioeng Transl Med 8:e10482 (2023). PubMed: 38023709