Recombinant Anti-Tissue Factor antibody [EPR22548-232] (ab228968)
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
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Tissue Factor antibody [EPR22548-232]
See all Tissue Factor primary antibodies -
Description
Rabbit monoclonal [EPR22548-232] to Tissue Factor -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type HAP1 whole cell lysate; A431, BxPC-3 and MDA-MB-231 whole cell lysate. IHC-P: Human cervix carcinoma and kidney tissue. ICC/IF: A431 cells. Flow Cyt: A431 cells. IP: A431 whole cell lysate.
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General notes
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.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22548-232 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab228968 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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WB |
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 33 kDa).
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IHC-P | (1) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/100.
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Flow Cyt |
1/500.
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 47 kDa (predicted molecular weight: 33 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
Flow Cyt
1/500. |
IP
1/30. |
Target
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Function
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. -
Sequence similarities
Belongs to the tissue factor family. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 2152 Human
- Omim: 134390 Human
- SwissProt: P13726 Human
- Unigene: 62192 Human
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Alternative names
- CD142 antibody
- CD142 antigen antibody
- Coagulation factor III (thromboplastin tissue factor) antibody
see all
Images
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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.
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All lanes : Anti-Tissue Factor antibody [EPR22548-232] (ab228968) at 1/1000 dilution
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.
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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. -
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.
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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 for detection (HRP) (ab131366), was used Detection Reagent 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.
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (6)
ab228968 has been referenced in 6 publications.
- Misasi R et al. Signal transduction pathway involved in platelet activation in immune thrombotic thrombocytopenia after COVID-19 vaccination. Haematologica 107:326-329 (2022). PubMed: 34615340
- Capozzi A et al. Effect of heparanase inhibitor on tissue factor overexpression in platelets and endothelial cells induced by anti-β2-GPI antibodies: Reply to comment from Mackman et al. J Thromb Haemost 20:261-262 (2022). PubMed: 34954878
- 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
- Li JC et al. Neutrophil extracellular traps participate in the development of cancer-associated thrombosis in patients with gastric cancer. World J Gastroenterol 28:3132-3149 (2022). PubMed: 36051331
- Zhang H et al. Tissue Factor-Enriched Neutrophil Extracellular Traps Promote Immunothrombosis and Disease Progression in Sepsis-Induced Lung Injury. Front Cell Infect Microbiol 11:677902 (2021). PubMed: 34336711
- Greimel T et al. Bile acid-induced tissue factor activity in hepatocytes correlates with activation of farnesoid X receptor. Lab Invest 101:1394-1402 (2021). PubMed: 34145381