Description
Tissue Factor (TF) is a 45 kDa transmembrane cell surface glycoprotein known for its role in initiating coagulation. It is comprised of three domains: an extracellular domain (aa 1-219), a hydrophilic spanning domain (aa 220-242) and a cytoplasmic tail (aa 243-263). Released into the blood stream following disruption of the endothelium, tissue factor functions as a receptor and cofactor for factors VII and VIIa. Contact between TF and blood is sufficient to initiate the extrinsic pathway of coagulation. This initiation requires the participation of a series of molecules; factor VII, factor X or factor IX, charged phospholipids and calcium ions, and their ability to complex with tissue factor. The TF/FVIIa complex efficiently activates both factor X and factor IX, thus initiating both the intrinsic and extrinsic coagulation pathways.

No. 4511 is highly inhibitory of rabbit tissue factor procoagulant activity (rabbit brain thromboplastin) in a single-stage prothrombin assay. It shows no effect on human tissue factor. While its epitope specificity is undetermined, its reactivity toward rabbit tissue factor in Western blot under non-reducing conditions and reduced reactivity toward rabbit tissue factor under reducing conditions suggests that the antibody recognizes an epitope that includes disulfide bonds.

Preparation
No. 4511 is a murine IgG1 monoclonal antibody purified from ascites fluid via Protein G affinity chromatography. Immunoaffinity purified rabbit tissue factor (Mr = 45,000) was used as the immunizing antigen.

Presentation
Screw-capped glass vial containing 100 μg of purified IgG1 lyophilized from a 200 μL solution of 0.15M Phosphate Buffered Saline, 100 mM Mannitol, pH 7.4.

Reconstitution
Add 200 μL of filtered deionized or sterile water to generate a 0.5 mg/mL stock solution.

Storage
Store lyophilized antibody at 2°-8°C. Aliquot and store reconstituted antibody at –20°C or colder.

Applications
A. Western Blot Analysis
No. 4511 at a primary antibody concentration of 5 μg/mL detects as little as 5 ng or native and recombinant rabbit tissue factor (ADI No. 4520) under non-reducing conditions. Multiple bands may be visualized due to variations in glycosylation.

B. Immunohistochemistry
No. 4511 has been used for localizing rabbit tissue factor in the adventitia of blood vessels and in normal and diseased rabbit glomeruli (paraffin-embedded tissue sections). A primary antibody concentration of 20 μg/mL in conjunction with Bouin’s fixation is recommended.1

C. Immunoprecipitation/Affinity Purification
No. 4511 is suitable for these applications. A 5:1 molar ration of antibody:protein is recommended for immunoprecipitation.

D. Inhibition
5 μg of No. 4511 inhibits 99% of the the procoagulant activity of rabbit tissue factor. It demonstrates no inhibitory effects toward human tissue factor.

References
1. Tipping, P. G., Erlich, J. H., Apostolopoulos, J., Mackman, N., Loskutoff, D. and Holdsworth, S. R. Glomerular tissue factor expression in crescentic glomerulonephritis. Correlations beween antigen, activity, and mRNA. American Journal of Pathology 1995, 147(6): 1736-1748.