Anti-Fibrinogen alpha chain antibody [EPR2919]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrinogen alpha chain antibody [EPR2919] (AB92572)

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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - 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.

• IP

Lab

Immunoprecipitation - Anti-Fibrinogen alpha chain antibody [EPR2919] (AB92572)

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.

All lanes:

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

Predicted band size: 95 kDa

Observed band size: 110 kDa

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Exposure time: 5s

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Fibrinogen alpha chain antibody [EPR2919] (AB92572)

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).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrinogen alpha chain antibody [EPR2919] (AB92572)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrinogen alpha chain antibody [EPR2919] (AB92572)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrinogen alpha chain antibody [EPR2919] (AB92572)

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.

• WB

Unknown

Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (AB92572)

All lanes:

Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572) at 1/10000 dilution

Lane 1:

Human plasma lysate at 15 µg

Lane 2:

Rat platelet lysate at 15 µg

Lane 3:

Rat plasma lysate at 15 µg

Secondary

All lanes:

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: 95 kDa

Observed band size: 95 kDa

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• WB

Supplier Data

Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (AB92572)

Blocking and diluting buffer : 5% NFDM/TBST.

The bands below 95kDa are probably FGA fragments (PMID : 24937656).

All lanes:

Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572) at 1/1000 dilution

All lanes:

Rat platelet tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 95 kDa

Observed band size: 95 kDa,35-60 kDa

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Exposure time: 20s

• WB

Lab

Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (AB92572)

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

All lanes:

Western blot - Anti-Fibrinogen alpha chain antibody [EPR2919] (ab92572) at 1/1000 dilution

Lane 1:

Mouse liver cell lysate at 20 µg

Lane 2:

Mouse plasma cell lysate at 20 µg

Lane 3:

Mouse platelet cell lysate at 20 µg

Secondary

All lanes:

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 1/100000 dilution

Predicted band size: 95 kDa

Observed band size: 87 kDa

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