Anti-Fibronectin antibody [EPR19241-46]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [EPR19241-46] (AB199056)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Hepa 1-6 (mouse hepatoma epithelial cell line) cells labeling Fibronectin with ab199056 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining on Hepa1-6 cells.

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

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR19241-46] (AB199056)

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Fibronectin with ab199056 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on the extracellular matrix of mouse stomach (PMID : 650151).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [EPR19241-46] (AB199056)

Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling Fironectin with ab199056 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on the extracellular matrix of rat stomach (PMID : 650151).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Fibronectin antibody [EPR19241-46] (AB199056)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Hepa1-6 (mouse hepatoma epithelial cell line) cell line labeling Fibronectin with ab199056 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluorr^® 488) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-Fibronectin antibody [EPR19241-46] (AB199056)

Fibronectin was immunoprecipitated from 0.35 mg of mouse plasma with ab199056 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab199056 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Mouse plasma 10 μg (Input).

Lane 2 : ab199056 IP in mouse plasma.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab199056 in mouse plasma.

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

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-Fibronectin antibody [EPR19241-46] (ab199056)

Predicted band size: 262 kDa

false

• WB

Supplier Data

Western blot - Anti-Fibronectin antibody [EPR19241-46] (AB199056)

Exposure time : Lanes 1 and 2 : 4 seconds; Lanes 3,4 and 5 : 2 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression profile and molecular weight observed is consistent with what has been described in the literature (PMID : 6994838, PMID 7516997, PMID : 21923916).

All lanes:

Western blot - Anti-Fibronectin antibody [EPR19241-46] (ab199056) at 1/1000 dilution

Lane 1:

Mouse liver lysate at 10 µL/mL Buffer

Lane 2:

Rat liver lysate at 10 µL/mL Buffer

Lane 3:

Mouse plasma at 10 µL/mL Buffer

Lane 4:

Rat plasma at 10 µL/mL Buffer

Lane 5:

Hepa1-6 (mouse hepatoma epithelial cell line) whole cell lysate at 10 µL/mL Buffer

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: 262 kDa

Observed band size: 220-273 kDa

true

• WB

CiteAb

Western blot - Anti-Fibronectin antibody [EPR19241-46] (AB199056)

Fibronectin western blot using anti-Fibronectin antibody [EPR19241-46] ab199056. Publication image and figure legend from Yang, G., Jin, L., et al., 2019, Mar Drugs, PubMed 31842414.

ab199056 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab199056 please see the product overview.

Fx can significantly reduce the accumulation of FN and collagen IV induced by HG in GMCs. (a) FN was detected by western blot after 24 h of HG treatment with or without Fx (2 μM), Rsv (40 μM) or MK2206 (2 μM). (b) After 24 h of HG treatment with or without Fx (2 μM), Rsv (40 μM) or MK2206 (2 μM), GMCs were lysed for protein extraction, and collagen IV was detected by western blot. (c) GMCs were induced by HG with or without Fx (2 μM), Rsv (40 μM), or MK2206 (2 μM) for 24 h. FN was stained by the immunofluorescence with antibody against FN and then captured under Olympus IX71 (400X); red fluorescence indicates FN, blue fluorescence indicates nucleus, and scale represents 10 μm. β-Actin was measured as a loading control; all the experiments were repeated at least three times independently with similar results. * p < 0.05 vs. normal glucose (NG) group and # p < 0.05 vs. 30 mM HG group.

false