Tissue factor pathway inhibitor (or TFPI) is a single-chain polypeptide which can reversibly inhibit factor Xa (Xa). While Xa is inhibited, the Xa-TFPI complex can subsequently also inhibit the FVIIa-tissue factor complex. TFPI contributes significantly to the inhibition of Xa in vivo, despite being present at concentrations of only 2.5 nM. TFPI has a relative molecular mass of 34,000 to 40,000 depending on the degree of proteolysis of the C-terminal region. TFPI consists of a highly negatively charged amino-terminus, three tandemly linked Kunitz domains, and a highly positively charged carboxy-terminus. With its Kunitz domains, TFPI exhibits significant homology with human inter-alpha-trypsin inhibitor and bovine basic pancreatic trypsin inhibitor.
Background References
1. Luo J et al. TFPI is a colonic crypt receptor for TcdB from hypervirulent clade 2 C. difficile. Cell. 2022 Mar
2. Xing M et al. TFPI inhibits breast cancer progression by suppressing ERK/p38 MAPK signaling pathway. Genes Genomics. 2022 Jul
Western blot analysis of TFPI on different lysates with Rabbit anti-TFPI antibody (HA722199) at 1/1,000 dilution.
Lane 1: HepG2 cell lysate Lane 2: LoVo cell lysate (negative) Lane 3: HUVEC cell lysate Lane 4: EA.hy926 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 35 kDa Observed band size: 35-45 kDa
Exposure time: 59 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722199) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
TFPI was immunoprecipitated from 0.2 mg HepG2 cell lysate with HA722199 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722199 at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HepG2 cell lysate (input) Lane 2: HA722199 IP in HepG2 cell lysate Lane 3: Rabbit IgG instead of HA722199 in HepG2 cell lysate
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-TFPI antibody (HA722199) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722199) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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