Western blot - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (ab207178)

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight is consistent with the literature.

Negative control: Human fetal liver (PMID: 10593948, PMID: 9139873).

All lanes:

Western blot - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (ab207178) at 1/1000 dilution

Lane 1:

WI-38 (Human fetal lung fibroblast cell line) whole cell lysate at 10 µg

Lane 2:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 10 µg

Lane 3:

Human fetal liver lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 88 kDa

Observed band size: 95 kDa

Exposure time: 3min

Western blot - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (ab207178)

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 1 minute; Lane 2: 3 minutes.

The molecular weight is consistent with the literature (PMID: 10593948).

All lanes:

Western blot - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (ab207178) at 1/1000 dilution

Lane 1:

HFF-1 (Human forskin fibroblast cell line) whole cell lysate at 10 µg

Lane 2:

IMR-90 (Human lung fibroblast cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 88 kDa

Observed band size: 95 kDa

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (ab207178)

Immunohistochemical analysis of paraffin-embedded human pancreatic cancer tissue labeling Fibroblast activation protein, alpha with ab207178 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cell surface staining on stromal cells of human pancreas cancer is observed [PMID:2371645].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

Western blot - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (ab207178)

Different batches of ab207178 were tested on IMR-90 (Human lung fibroblast cell line) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 95 kDa.

All lanes:

Western blot - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (ab207178)

Predicted band size: 88 kDa

Observed band size: 95 kDa