Anti-Fibronectin 抗体 [F1]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [F1] (AB32419)

ICC/IF image of unpurified ab32419 stained HepG2 (Human liver hepatocellular carcinoma cell line) cells. The cells were fixed in 4% formaldehyde (10 minutes) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab32419 at 1/100 dilution overnight at +4°C. The secondary antibody (green) was DyLight^® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1 hour. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 μM.

Alexa Fluor^® 488 (ab198933) and Alexa Fluor^® 647 (ab198934) conjugated versions are available for this clone.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Fibronectin antibody [F1] (AB32419)

ICC/IF image of unpurified ab32419 stained human mesenchymal stem cells. The cells were fixed in paraformaldehyde and then incubated in 0.1%BSA / 1% goat serum for 30 minutes, to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32419, 1/100 dilution) for 2 hours at 22°C. The secondary antibody (green) was Alexa Fluor^® 488 goat anti-rabbit IgG. DAPI was used to stain the cell nuclei (blue).

This image is courtesy of an anonymous Abreview.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Fibronectin antibody [F1] (AB32419)

Intracellular Flow Cytometry analysis of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Fibronectin with purified ab32419 at 1/20 dilution (10μg/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr^®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibronectin antibody [F1] (AB32419)

Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab32419 at a dilution of 1/250. The secondary antibody used is ab97051 a HRP-conjugated goat anti-rabbit IgG (H+L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

• WB

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Western blot - Anti-Fibronectin antibody [F1] (AB32419)

Blocking/Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Fibronectin antibody [F1] (ab32419) at 1/5000 dilution

Lane 1:

Human serum at 20 µg

Lane 2:

Human plasma at 20 µg

Lane 3:

SK-OV-3 (Human ovarian cancer cell line) cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 262 kDa

Observed band size: 263 kDa

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• WB

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Western blot - Anti-Fibronectin antibody [F1] (AB32419)

Blocking/Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Fibronectin antibody [F1] (ab32419) at 1/1000 dilution

All lanes:

HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 262 kDa

Observed band size: 263 kDa

false