Recombinant Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Fibrinogen alpha chain with purified ab92572 at 1/50 dilution (3.16 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Intracellular Intracellular Flow Cytometry analysis of Hepa1-6 (Mouse hepatoma epithelial cell) labeling Fibrinogen alpha chain with ab92572 at 1/50 (red). Goat anti rabbit IgG (Alexa Fluor^® 488), ab150077 at 1/2000 was used as the secondary antibody. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control / colour Rabbit monoclonal IgG (ab172730) (black). Unlabelled control / colour Cell without incubation with primary antibody and secondary antibody (blue).

The band around 35 kD represents FGA fragment (PMID: 24937656).

Exposure time:
Lane 1: 3 minutes;
Lanes 2-3: 5.5 seconds

Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST

Immunohistochemical analysis of paraffin-embedded mouse placenta using ab92572 at 1/1000 dilution (0.5 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. A rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody (ready to use). Negative control used only the secondary antibody. Counterstained with haematoxylin.

Positive staining in plasma of mouse placenta.

The section was incubated with ab92572 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Immunohistochemical analysis of paraffin-embedded mouse lung using ab92572 at 1/1000 dilution (0.5 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins, a rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody (ready to use). Negative control used only the secondary antibody. Counterstained with haematoxylin.

Strong positive staining in plasma of mouse lung (PMID: 26259584).

The section was incubated with ab92572 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

ab92572 at a dilution of 1/100 immunoprecipitating Fibrinogen alpha chain in human plasma.

Lane 1 (input): Human plasma (10 µg)
Lane 2 (+): ab92572 + human plasma.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92572 in human plasma.

For western blotting, ab92572 was used as the primary antibody at a 1/1000 dilution and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Blocking and dilution buffer: 5% NFDM/TBST.