Anti-Prothrombin antibody [EPR20159] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Prothrombin antibody [EPR20159] - BSA and Azide free (AB251496)

This data was developed using ab208589, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Prothrombin with ab208589 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Prothrombin antibody [EPR20159] - BSA and Azide free (AB251496)

This data was developed using ab208589, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Prothrombin with ab208589 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IP

Lab

Immunoprecipitation - Anti-Prothrombin antibody [EPR20159] - BSA and Azide free (AB251496)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208589).

Prothrombin was immunoprecipitated from HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab208589 at 1/50 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab208589 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Prothrombin antibody [EPR20159] (<a href='/en-us/products/primary-antibodies/prothrombin-antibody-epr20159-ab208589'>ab208589</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate (Input) at 10 µg

Lane 2:

HepG2 whole cell lysate (+)

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/prothrombin-antibody-epr20159-ab208589'>ab208589</a> in HepG2 whole cell lysate (-)

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 33-72 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Prothrombin antibody [EPR20159] - BSA and Azide free (AB251496)

This data was developed using ab208589, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Prothrombin is proteolytically cleaved into different fragments. The fragments profile is consistent with what has been described in the literature (PMID : 17637839; 21131592; 16734589).

All lanes:

Western blot - Anti-Prothrombin antibody [EPR20159] (<a href='/en-us/products/primary-antibodies/prothrombin-antibody-epr20159-ab208589'>ab208589</a>) at 1/1000 dilution

Lane 1:

Human plasma at 20 µg

Lane 2:

Human fetal liver tissue lysate at 20 µg

Lane 3:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/4000 dilution

Predicted band size: 70 kDa

Observed band size: 33 kDa,52 kDa,72 kDa

false

Exposure time: 3min