Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, ELISA Capture (Matched Antibody Pair), Immunoprecipitation

Cited:

Immunocytochemistry, ELISA Development

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 323514
View all Coagulation Factor III/Tissue Factor Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Coagulation Factor III
Gly34-Glu251
Accession # P13726

Specificity

Detects human coagulation factor III/Tissue Factor in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant mouse Tissue Factor is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human Coagulation Factor III/Tissue Factor Antibody

Detection of Human Coagulation Factor III antibody by Western Blot.

Detection of Human Coagulation Factor III by Western Blot.

Western blot shows lysates of THP-1 human acute monocytic leukemia cell line treated (+) with TNF-a. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Coagulation Factor III/Tissue Factor Monoclonal Antibody (Catalog # MAB2339) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Coagulation Factor III at approximately 50 and 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Coagulation Factor III/Tissue Factor by Western Blot.

Western blot shows lysates of A431 human epithelial carcinoma cell line and A549 human lung carcinoma cell line (negative control). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Coagulation Factor III/Tissue Factor Monoclonal Antibody (Catalog # MAB2339) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Coagulation Factor III/Tissue Factor at approximately 48 kDa (as indicated). GAPDH (MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
Detection of Coagulation Factor III/Tissue Factor by Western Blot

Detection of Coagulation Factor III/Tissue Factor by Western Blot

SC1 inhibits TF-dependent tumor cell migration(A-C) MDA-MB-231 and BxPC3 cells transfected with TRIPZ non-targeting ShRNA (Sh-NT) or TF ShRNA (Sh-TF) were induced with doxycycline (Dox) for 5-7 days to deplete TF. The cells were subjected to immunoblotting (A) or cell migration assay. Migrated cells were stained with crystal violet (B) and the quantified results are plotted (C). (D-F) The indicated cell lines were assayed similarly for cell migration with various doses of SC1 or IgG (D, E) or 100 nM SC1 (F). Quantified results are plotted. *, P<0.05; **, P<0.01; ***, P<0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28938620), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Coagulation Factor III/Tissue Factor by Western Blot

Detection of Coagulation Factor III/Tissue Factor by Western Blot

Mechanistic target of rapamycin (mTOR) inhibitor differentially targets TF-mRNA transcription or TF protein degradation in a cellular context-dependent manner. (A,B) U87MG and HCC827 (A), H1975 and PC9 (B) cells were treated as indicated, subjected to qPCR analysis of TF mRNA as described in section “Materials and Methods.” The relative mRNA levels are shown. (C,D) H1975 (C) and PC9 (D) cells were treated with 5 μmol/L MTI-31 alone or in combination with 10 μmol/L CQ or 0.01 μmol/L PS-341 for 24 h, then immunoblotted (left panels) and quantified (right panels). *P < 0.05; **P < 0.01, ***P < 0.001. (E) The amino acid sequence of TF was download from https://www.ncbi.nlm.nih.gov/to set up the model of TF structure using Pymol software. The KFERQ-motif is marked in yellow and its sequence is marked in red. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32923403), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Coagulation Factor III/Tissue Factor by Western Blot

Detection of Coagulation Factor III/Tissue Factor by Western Blot

Mechanistic target of rapamycin (mTOR) promotes TF overexpression in EGFR-mut tumors. (A) mTOR inhibitor WYE-132 and AZD8055 decreased TF expression in four indicated cell lines. The indicated cells were treated for 24 h in full growth medium followed by immunoblotting. (B) The indicated cells were serum-starved then stimulated with 50 ng/mL EGF without or with inhibitor for 24 h. (C) Serum-starved U87MG cells were stimulated with 10% serum without or with WYE-132 for 12 h and immunoblotted. (D,E) The indicated cell lines were infected with pGIPZ-shRNA encoding lentivirus, incubated for 6–7 days in full-growth medium and immunoblotted. (F) Cells as in D and E were serum-starved, stimulated with EGF and immunoblotted. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32923403), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Coagulation Factor III/Tissue Factor by Western Blot

Detection of Coagulation Factor III/Tissue Factor by Western Blot

Mechanistic target of rapamycin (mTOR) inhibitor differentially targets TF-mRNA transcription or TF protein degradation in a cellular context-dependent manner. (A,B) U87MG and HCC827 (A), H1975 and PC9 (B) cells were treated as indicated, subjected to qPCR analysis of TF mRNA as described in section “Materials and Methods.” The relative mRNA levels are shown. (C,D) H1975 (C) and PC9 (D) cells were treated with 5 μmol/L MTI-31 alone or in combination with 10 μmol/L CQ or 0.01 μmol/L PS-341 for 24 h, then immunoblotted (left panels) and quantified (right panels). *P < 0.05; **P < 0.01, ***P < 0.001. (E) The amino acid sequence of TF was download from https://www.ncbi.nlm.nih.gov/to set up the model of TF structure using Pymol software. The KFERQ-motif is marked in yellow and its sequence is marked in red. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32923403), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Coagulation Factor III/Tissue Factor by Western Blot

Detection of Coagulation Factor III/Tissue Factor by Western Blot

Mechanistic target of rapamycin (mTOR) promotes TF overexpression in EGFR-mut tumors. (A) mTOR inhibitor WYE-132 and AZD8055 decreased TF expression in four indicated cell lines. The indicated cells were treated for 24 h in full growth medium followed by immunoblotting. (B) The indicated cells were serum-starved then stimulated with 50 ng/mL EGF without or with inhibitor for 24 h. (C) Serum-starved U87MG cells were stimulated with 10% serum without or with WYE-132 for 12 h and immunoblotted. (D,E) The indicated cell lines were infected with pGIPZ-shRNA encoding lentivirus, incubated for 6–7 days in full-growth medium and immunoblotted. (F) Cells as in D and E were serum-starved, stimulated with EGF and immunoblotted. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32923403), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Coagulation Factor III/Tissue Factor by Western Blot

Detection of Coagulation Factor III/Tissue Factor by Western Blot

Mechanistic target of rapamycin (mTOR) promotes TF overexpression in EGFR-mut tumors. (A) mTOR inhibitor WYE-132 and AZD8055 decreased TF expression in four indicated cell lines. The indicated cells were treated for 24 h in full growth medium followed by immunoblotting. (B) The indicated cells were serum-starved then stimulated with 50 ng/mL EGF without or with inhibitor for 24 h. (C) Serum-starved U87MG cells were stimulated with 10% serum without or with WYE-132 for 12 h and immunoblotted. (D,E) The indicated cell lines were infected with pGIPZ-shRNA encoding lentivirus, incubated for 6–7 days in full-growth medium and immunoblotted. (F) Cells as in D and E were serum-starved, stimulated with EGF and immunoblotted. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32923403), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Coagulation Factor III/Tissue Factor Antibody

Application
Recommended Usage

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Human Coagulation Factor III/Tissue Factor (Catalog # 2339-PA), see our available Western blot detection antibodies

Western Blot

1-2 µg/mL
Sample: THP‑1 human acute monocytic leukemia cell line treated with TNF-alpha and A431 human epithelial carcinoma cell line

Human Coagulation Factor III/Tissue Factor Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human Coagulation Factor III/ Tissue Factor Biotinylated Antibody (Catalog # BAF2339)
  • Standard: Recombinant Human Coagulation Factor III/Tissue Factor, CF (Catalog # 2339-PA)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using MAB2339 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Coagulation Factor III/Tissue Factor

Coagulation Factor III/Tissue Factor (TF), also known as thromboplastin and CD142, is a type I transmembrane protein found in a variety of cell types. It functions as a protein cofactor/receptor of Coagulation Factor VII, which is synthesized in the liver and circulated in the plasma (1). Upon binding of TF, the inactive factor VII is rapidly converted into activated VIIa. The resulting 1:1 complex of VIIa and TF initiates the coagulation pathway and has also important coagulation-independent functions such as angiogenesis (2). TF is synthesized as a 295 amino acid (aa) precursor,  with a signal peptide (aa 1-32), an extracellular domain (aa 33-251), a transmembrane region (aa 252-274) and a cytoplasmic tail (aa 275-295) (3‑6).

References

  1. Morrissey, J.H. (2004) in Handbook of Proteolytic Enzymes. Barrett, A.J. et al. (ed) San Diego, Academic Press, p. 1659. 
  2. Versteeg, H.H. et al. (2003) Carcinogenesis 24:1009. 
  3. Scarpati, E.M. et al. (1987) Biochemistry 26:5234.
  4. Fisher, K.L. et al. (1987) Thromb. Res. 48:89.
  5. Morrissey, J.H. et al. (1987) Cell 50:129.
  6. Spicer, E.K. (1987) Proc. Natl. Acad. Sci. USA 84:5148.

Alternate Names

CD142, F3, Thromboplastin, Tissue Factor

Entrez Gene IDs

2152 (Human); 14066 (Mouse)

Gene Symbol

F3

UniProt

P13726

Additional Coagulation Factor III/Tissue Factor Products

Product Documents for Human Coagulation Factor III/Tissue Factor Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Coagulation Factor III/Tissue Factor Antibody

For research use only

Citations for Human Coagulation Factor III/Tissue Factor Antibody

Customer Reviews for Human Coagulation Factor III/Tissue Factor Antibody (2)

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  • Human Coagulation Factor III/Tissue Factor Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 07/06/2021
    Human Coagulation Factor III/Tissue Factor Antibody MAB2339
  • Human Coagulation Factor III/Tissue Factor Antibody
    Name: Zoia Levashova
    Application: Meso Scale Discovery
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 06/22/2018
    After biotinylation, used as a capture reagent according to the manufacturer’s protocol (Meso Scale Diagnostics LLC). Dynamic range for human TF was 10-25,000 pg/mL.
    Human Coagulation Factor III/Tissue Factor Antibody MAB2339

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Associated Pathways

Blood Coagulation Signaling Pathways Blood Coagulation Signaling Pathway Thumbnail