Anti-Fibroblast activation protein, alpha 抗体 [EPR20021]

• IHC-P

AbReview62299****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (AB207178)

Formalin-fixed paraffin-embedded human colon cancer tissue labeling Fibroblast activation protein alpha using ab207178 at 1/100 dilution in immunohistochemical analysis. Heat mediated antigen retieval using Citrate pH 6.0.

Courtesy of an anonymous abreview

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (AB207178)

Immunohistochemical analysis of formalin fixed paraffin embedded human colon carcinoma labelling Fibroblast activation protein, alpha with ab207178 at a concentration of 3 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab207178 anti- Fibroblast activation protein, alpha [EPR20021] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• IHC-P

Supplier Data

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (AB207178)

Immunohistochemical analysis of paraffin-embedded human liver 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.

Negative control : no staining on human liver [PMID : 9139873].

Counter stained with Hematoxylin.

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

• IHC-P

AbReview57315****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (AB207178)

Immunohistochemical analysis of formaldehyde fixed paraffin embedded human Ovarian carcinoma tissue sectioins labeling Fibroblast activation protein with ab207178 at 1/500. Biotin conjugated goat anti-rabbit IgG at 1/300 was used as the secondary antibody. Antigen retrieval was heat mediated using citric acid.

This image is of an Abreciew submitted byCarl Hobbs.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (AB207178)

Immunohistochemical analysis of paraffin-embedded Human stomach cancer tissue labeling Fibroblast activation protein, alpha with ab207178 at 1/1000 (0.718 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on stromal fibroblast cells of human stomach cancer. The section was incubated with ab207178 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-Fibroblast activation protein, alpha antibody [EPR20021] (AB207178)

Immunohistochemical analysis of paraffin-embedded human pancreas 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.

Negative control : no staining on human pancreas [PMID : 22371645].

Counter stained with Hematoxylin.

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-Fibroblast activation protein, alpha antibody [EPR20021] (AB207178)

Immunohistochemical analysis of paraffin-embedded human breast 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 breast cancer is observed [PMID : 18628473].

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (AB207178)

Immunohistochemical analysis of paraffin-embedded Human sciatic nerve tissue labeling Fibroblast activation protein, alpha with ab207178 at 1/1000 (0.718 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human sciatic nerve. The section was incubated with ab207178 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.

• WB

Supplier Data

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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 88 kDa

Observed band size: 95 kDa

false

• WB

Lab

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

false

• WB

Lab

Western blot - Anti-Fibroblast activation protein, alpha antibody [EPR20021] (AB207178)

Western blot : Anti-Fibroblast activation protein, alpha antibody [EPR20021] ab207178 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 80-100 kDa. 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-Fibroblast activation protein, alpha antibody [EPR20021] (ab207178) at 1/1000 dilution

Lane 1:

Wild-type MCF7 BFA (5 g/mL, 6h) at 20 µg

Lane 2:

Wild-type MCF7 BFA (0 g/mL, 6h) at 20 µg

Lane 3:

FAP knockout MCF7 BFA (5 g/mL, 6 h) at 20 µg

Lane 4:

FAP knockout MCF7 BFA (0 g/mL, 6 h) at 20 µg

Secondary

All lanes:

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

Predicted band size: 80-100 kDa

Observed band size: 80-100 kDa

false

• WB

Supplier Data

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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 88 kDa

Observed band size: 95 kDa

false

Exposure time: 3min