Anti-TFPI antibody [EPR22977-16]

• Flow Cyt

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Flow Cytometry - Anti-TFPI antibody [EPR22977-16] (AB260041)

Flow cytometric analysis of HepG2 (human hepatocellular carcinoma epithelial cell) cells labeling TFPI with ab260041 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• IP

Unknown

Immunoprecipitation - Anti-TFPI antibody [EPR22977-16] (AB260041)

TFPI was immunoprecipitated from 0.35 mg human placenta lysate 10μg with ab260041 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab260041 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Human placenta lysate 10μg

Lane 2 : ab260041 IP in human placenta lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab260041 in human placenta lysate

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-TFPI antibody [EPR22977-16] (ab260041)

Predicted band size: 35 kDa

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• WB

Lab

Western blot - Anti-TFPI antibody [EPR22977-16] (AB260041)

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 secs; Lane 3 : 3 secs; Lanes 4-5 : 15 secs; Lanes 6-7 : 59 secs.

All lanes:

Western blot - Anti-TFPI antibody [EPR22977-16] (ab260041) 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:

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

Lane 6:

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

Lane 7:

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 - 7:

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

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