重组Anti-Tissue Factor抗体[EPR22548-232] - BSA and Azide free (ab254010)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22548-232] to Tissue Factor - BSA and Azide free
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Tissue Factor抗体[EPR22548-232] - BSA and Azide free
参阅全部 Tissue Factor 一抗 -
描述
兔单克隆抗体[EPR22548-232] to组织Factor - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details -
种属反应性
与反应: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- IHC-P: Human cervix carcinoma and kidney tissue. ICC/IF: A431 cells. Flow Cyt: A431 cells. IP: Tissue Factor IP in A431 whole cell lysate.
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常规说明
ab254010 is the carrier-free version of ab228968.
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.2
Constituent: 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
EPR22548-232 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Immunohistochemistry kits
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab254010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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WB |
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 33 kDa).
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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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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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说明 |
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WB
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 33 kDa). |
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. |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
靶标
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功能
Initiates blood coagulation by forming a complex with circulating factor VII or VIIa. The [TF:VIIa] complex activates factors IX or X by specific limited protolysis. TF plays a role in normal hemostasis by initiating the cell-surface assembly and propagation of the coagulation protease cascade. -
序列相似性
Belongs to the tissue factor family. -
细胞定位
Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 2152 Human
- Omim: 134390 Human
- SwissProt: P13726 Human
- Unigene: 62192 Human
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别名
- CD142 antibody
- CD142 antigen antibody
- Coagulation factor III (thromboplastin tissue factor) antibody
see all
图片
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Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Tissue Factor with ab228968 at 1/500 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in human renal glomerulus (PMID: 7684196). The section was incubated with ab228968 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228968).
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All lanes :
Lane 1 : Wild-type HAP1 whole cell lysate at 60 µg
Lane 2 : Tissue Factor knockout HAP1 whole cell lysate at 60 µg
Lane 3 : A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 20 µg
Lane 4 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 5 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 6 : BxPC-3 (human pancreas adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 33 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?ab228968 was shown to specifically react with Tissue Factor in wild-type HAP1 cells as signal was lost in Tissue Factor knockout cells. Wild-type and Tissue Factor knockout samples were subjected to SDS-PAGE. ab228968 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.
The molecular weight observed is consistent with what has been described in the literature (PMID: 28938620).
Low expression: MCF-7 (PMID: 24137414, 28938620).
Exposure times: Lanes 1-2: 70 secs; Lanes 3-6: 3.25 secs.
Blocking/Dilution buffer: 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228968).
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Immunohistochemical analysis of paraffin-embedded human cervix carcinoma tissue labeling Tissue Factor with ab228968 at 1/500 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous and weak cytoplasmic staining on tumor cells of human cervix carcinoma (PMID: 26383146). The section was incubated with ab228968 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Perform heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilizedA431 (human epidermoid carcinoma epithelial cell) and MCF7 () cells labeling Tissue Factor with ab228968 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in A431 cell line. Low expression: MCF7 PMID: 24137414, 28938620. The nuclear stain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228968).
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Flow cytometric analysis of MCF7 (human breast adenocarcinoma epithelial cell, Left) / A431 (human epidermoid carcinoma epithelial cell, Right) labeling Tissue Factor with ab228968 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Low expression: MCF7 (PMID: 24137414, PMID: 28938620).
Gated on viable cells.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228968).
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Tissue Factor was immunoprecipitated from 0.35 mg A431 (human epidermoid carcinoma epithelial cell) whole cell lysate with ab228968 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab228968 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: A431 whole cell lysate 10 µg (Input).
Lane 2: ab228968 IP in A431 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab228968 in A431 whole cell lysate.Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 30 seconds.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab228968).
实验方案
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.
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (1)
ab254010 被引用在 1 文献中.
- Mackman N et al. Effect of heparanase inhibitor on tissue factor overexpression in platelets and endothelial cells induced by anti-β2-GPI antibodies: Comment from Mackman et al. J Thromb Haemost 20:260-261 (2022). PubMed: 34954879