Anti-TFPI antibody [EPR22977-133]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-TFPI antibody [EPR22977-133] (AB260042)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HUVEC (human umbilical vein endothelial cell) cells labeling TFPI with ab260042 at 1/100 (6.8 μg/ml) dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 (2 μg/ml) dilution (Green). Confocal image showing cytoplasmic staining in HUVEC cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-TFPI antibody [EPR22977-133] (AB260042)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling TFPI with ab260042 at 1/100 (6.8 μg/ml) dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 (2 μg/ml) dilution (Green). Confocal image showing cytoplasmic and membranous staining in HepG2 cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

• WB

Lab

Western blot - Anti-TFPI antibody [EPR22977-133] (AB260042)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 22144186).

Negative control : Human colon and LoVo (PMID : 20062932, 9311608).

Exposure times : Lanes 1-2 : 59 seconds; Lane 3 : 26 seconds; Lanes 4-8 : 59 seconds.

All lanes:

Western blot - Anti-TFPI antibody [EPR22977-133] (ab260042) at 1/1000 dilution

Lane 1:

Human placenta lysate at 20 µg

Lane 2:

Human colon lysate at 20 µg

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

HUVEC (human umbilical vein endothelial cell) whole cell lysate at 20 µg

Lane 7:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 8:

LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Lanes 3 - 8:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 35 kDa

Observed band size: 45 kDa,47 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-TFPI antibody [EPR22977-133] (AB260042)

TFPI was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10μg with ab260042 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab260042 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Lane 1 : HepG2 whole cell lysate 10μg

Lane 2 : ab260042 IP in HepG2 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab260042 in HepG2 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-TFPI antibody [EPR22977-133] (ab260042)

Predicted band size: 35 kDa

false