Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue sections labeling Fibrinogen alpha chain with purified ab92572 at 1/3000 dilution (0.053 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

All lanes : Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572) at 1/10000 dilution (purified)

Lane 1 : Human plasma lysate
Lane 2 : Rat platelet lysate
Lane 3 : Rat plasma lysate

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 95 kDa
Observed band size: 95 kDa

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat placenta tissue sections labeling Fibrinogen alpha chain with purified ab92572 at 1/3000 dilution (0.053 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

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.

 

All lanes : Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572) at 1/1000 dilution

Lane 1 : Mouse liver cell lysate
Lane 2 : Mouse plasma cell lysate
Lane 3 : Mouse platelet cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 95 kDa
Observed band size: 87 kDa why is the actual band size different from the predicted?

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.