Anti-Fibrinogen gamma chain antibody [EP24539-127]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrinogen gamma chain antibody [EP24539-127] (AB281924)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labelling Fibrinogen gamma chain with ab281924 at 1/4000 (0.135 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in the blood plasma and stroma of human pancreas (PMID : 16510577). The section was incubated with ab281924 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrinogen gamma chain antibody [EP24539-127] (AB281924)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labelling Fibrinogen gamma chain with ab281924 at 1/4000 (0.135 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human hepatocellular carcinoma (PMID : 30863175). The section was incubated with ab281924 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Fibrinogen gamma chain antibody [EP24539-127] (AB281924)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HepG2 cells labelling Fibrinogen gamma chain with ab281924 at 1/100 (5.38 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green) Confocal image showing cytoplasmic staining in HepG2 cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibrinogen gamma chain antibody [EP24539-127] (AB281924)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling Fibrinogen gamma chain with ab281924 at 1/4000 (0.135 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human liver (PMID : 24648543). The section was incubated with ab281924 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-Fibrinogen gamma chain antibody [EP24539-127] (AB281924)

Fibrinogen gamma chain was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug with ab281924 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab281924 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug.

Lane 2 : ab281924 IP in HepG2 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab281924 in HepG2 whole cell lysate.

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-Fibrinogen gamma chain antibody [EP24539-127] (ab281924)

Predicted band size: 52 kDa

Observed band size: 50 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Fibrinogen gamma chain antibody [EP24539-127] (AB281924)

Fibrinogen gamma chain was immunoprecipitated from 0.35 mg Mouse plasma lysate 10 ug with ab281924 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab281924 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse plasma lysate 10 ug.

Lane 2 : ab281924 IP in Mouse plasma lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab281924 in mouse plasma lysate.

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-Fibrinogen gamma chain antibody [EP24539-127] (ab281924)

Predicted band size: 52 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-Fibrinogen gamma chain antibody [EP24539-127] (AB281924)

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

Exposure time : Lanes 1 : 1 seconds.

Lane 2 : 26 seconds.

All lanes:

Western blot - Anti-Fibrinogen gamma chain antibody [EP24539-127] (ab281924) at 1/1000 dilution

Lane 1:

Human liver lysate at 10 µg

Lane 2:

Mouse liver lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Predicted band size: 52 kDa

Observed band size: 100 kDa,50 kDa

false

• WB

Lab

Western blot - Anti-Fibrinogen gamma chain antibody [EP24539-127] (AB281924)

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

Low expression : A549/DU145 (PMID : 17949478).

All lanes:

Western blot - Anti-Fibrinogen gamma chain antibody [EP24539-127] (ab281924) at 1/1000 dilution

Lane 1:

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

Lane 2:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 3:

DU145 (human prostate carcinoma epithelial cell), whole cell lysate at 10 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 1/50000 dilution

Predicted band size: 52 kDa

Observed band size: 50 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-Fibrinogen gamma chain antibody [EP24539-127] (AB281924)

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 : 21480334).

Exposure time : Lanes 1-5 : 10 seconds

Lane 6 : 37 seconds.

All lanes:

Western blot - Anti-Fibrinogen gamma chain antibody [EP24539-127] (ab281924) at 1/5000 dilution

Lane 1:

Human plasma lysate at 10 µg

Lane 2:

Human plasma lysate treated with PNGase F whole cell lysate at 10 µg

Lane 3:

Mouse plasma lysate at 10 µg

Lane 4:

Mouse plasma lysate treated with PNGase F whole cell lysate at 10 µg

Lane 5:

Rat plasma lysate at 10 µg

Lane 6:

Rat liver lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 100 kDa,50 kDa

false