rabbit polyclonal anti tfpi antibody  (Cell Signaling Technology Inc)

Journal: Research and Practice in Thrombosis and Haemostasis

Article Title: Evaluation of a new commercial assay for the measurement of tissue factor activity of extracellular vesicles isolated from human plasma

doi: 10.1016/j.rpth.2025.103007

Figure Lengend Snippet: Measurement of extracellular vesicle (EV) tissue factor (TF) activity using the CY-QUANT MV-TF (CY) and Chapel Hill (CH) assays in the presence of an anti-TF pathway inhibitor (TFPI) antibody and levels of TFPI in EVs. Plasma was prepared from the whole blood of 4 healthy donors with lipopolysaccharide (+LPS) or without lipopolysaccharide (−LPS) stimulation. EVs were isolated from plasma by centrifugation at 20,000 × g for 60 minutes at 4 °C. (A) TF-dependent activated factor X generation was measured using the CY (yellow) and CH (blue) assays in the presence of an inhibitory anti-TFPI antibody (2H8). (B) TFPI was analyzed in EV samples from −LPS and +LPS samples by Western blotting. A polyclonal rabbit anti-human TFPI antibody was used (final concentration 3 μg/mL) to detect TFPI. A goat anti-rabbit immunoglobulin G secondary antibody (Cell Signaling, Cat. number 7074) was used at a 1:5000 dilution to detect the anti-TFPI antibody. The position of TFPI and molecular weight (MW) standards is shown. (C) Bands were quantified using ImageLab software. The Wilcoxon matched-pairs signed-rank test was used to compare the 2 groups. Data are shown as mean ± SD or individual values. ∗∗ P < .01.

Article Snippet: The membranes were incubated with a rabbit polyclonal anti-TFPI antibody [ ] (final concentration 3 μg/mL in 5% bovine serum albumin in 1× Tris-buffered saline with 0.1% Tween-20) overnight at 4 °C with rocking, washed, and then incubated with a goat anti-rabbit IgG antibody conjugated to horseradish peroxidase (Cell Signaling, Cat. number 7074, 1:5000 in blocking buffer) for 1 hour at room temperature with rocking.

Techniques: Activity Assay, Clinical Proteomics, Isolation, Centrifugation, Western Blot, Concentration Assay, Molecular Weight, Software