Anti-FAP antibody [RM1080]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FAP antibody [RM1080] (AB314456)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling FAP with ab314456 at 1/1000 (0.537 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human liver. The section was incubated with ab314456 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 was performed 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-FAP antibody [RM1080] (AB314456)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling FAP with ab314456 at 1/1000 (0.537 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the stroma in human lung carcinoma. The section was incubated with ab314456 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 was performed 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-FAP antibody [RM1080] (AB314456)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labeling FAP with ab314456 at 1/1000 (0.537 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the stroma in human colon carcinoma. The section was incubated with ab314456 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FAP antibody [RM1080] (AB314456)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized WI-38 (human fetal lung fibroblast cell) cells labelling FAP with ab314456 at 1/100 (5.37 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic staining in WI-38 cell line. Negative control : Jurkat.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• WB

Lab

Western blot - Anti-FAP antibody [RM1080] (AB314456)

Western blot : Rabbit Recombinant Multiclonal[RM1080] to FAP ab314456 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 90-100 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in FAP knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-FAP antibody [RM1080] (ab314456) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG at 20 µg

Lane 2:

Western blot - Human FAP knockout U-87 MG cell line (ab306819) at 20 µg

Lane 3:

WI-38 at 20 µg

Lane 4:

A549 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 88 kDa

Observed band size: 90-100 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-FAP antibody [RM1080] (AB314456)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : Jurkat. Samples are non-boiled as boiling may cause protein aggregation. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-FAP antibody [RM1080] (ab314456) at 1/1000 dilution

Lane 1:

HFF-1 (human skin fibroblast) whole cell lysate at 20 µg

Lane 2:

IMR-90 (human lung fibroblast) whole cell lysate at 20 µg

Lane 3:

WI-38 (human fetal lung fibroblast) whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole 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 1/20000 dilution

Observed band size: 95 kDa

false

Exposure time: 37s