All lanes : Anti-STAT1 antibody [EPRR21057-168] (ab210524) at 1/1000 dilution

Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2 : HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 5 : Human breast cancer lysate at 10 µg
Lane 6 : Human colon cancer lysate at 10 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Predicted band size: 87 kDa

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

Exposure times: Lane 1, 2 and 5: 3 minutes; Lane 3: 84 seconds; Lane 4 and 6: 32 seconds.

The doublet observed in some lanes likely represent the α and β isoforms of STAT1 (PMID: 8647800).

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling STAT1 with ab210524 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining mainly on interfollicular cells of human tonsil (PMID: 25336386; PMID: 25921060) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized MCF7 (Human breast adenocarcinoma cell line) cell line labeling STAT1 with ab210524 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

STAT1 was immunoprecipitated from 0.35 mg MCF7 (Human breast adenocarcinoma cell line) whole cell lysate with ab210524 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab210524 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: MCF7 whole cell lysate 10 µg (Input).
Lane 2: ab210524 IP in MCF7 whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab210524 in MCF7 whole cell lysate (-).

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 15 seconds.

The doublet observed in some lanes likely represent the α and β isoforms of STAT1 (PMID: 8647800).

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cell line labeling STAT1 with ab210524 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

Immunohistochemical analysis of paraffin-embedded paired human endometrial cancer and non-tumor endometrium tissue labeling STAT1 with ab210524 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Much higher staining intensity of endometrial cancer (Panel A) than its paired non-tumor endometrium (Panel B) (PMID: 25267067) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling STAT1 with ab210524 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining on glomerulus and some renal tubules of human kidney (PMID: 26678048) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).