Anti-ROS1 antibody [EPMGHR2]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROS1 antibody [EPMGHR2] (AB189925)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling ROS1 with Purified ab189925 at 1 : 300 dilution (5.23 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH9.0. Tissue was counterstained with hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROS1 antibody [EPMGHR2] (AB189925)

Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling ROS1 with unpurified ab189925 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with hematoxylin. (Inset : negative control).

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

• WB

Lab

Western blot - Anti-ROS1 antibody [EPMGHR2] (AB189925)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM /TBST We advise using U-118MG (FIG-ROS1 fusion) or HCC78 (SLC34A2-ROS1 fusion) as a positive cell line by WB. We have been unable to identify a positive control cell line for endogenous full-length ROS1. The identity of the lower MW band at approximately 60kDa is unknown.

All lanes:

Western blot - Anti-ROS1 antibody [EPMGHR2] (ab189925) at 1/1000 dilution

Lane 1:

U-118 MG (Human brain glioblastoma) whole cell lysate at 20 µg

Lane 2:

Saos-2 (Human osteosarcoma epithelial) whole cell lysate at 20 µg

Lane 3:

SW1353 (Human chondrosarcoma fibroblast) 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

Predicted band size: 264 kDa

Observed band size: 100 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-ROS1 antibody [EPMGHR2] (AB189925)

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

U-118MG cells carry the FIG-ROS1 fusion (PMID : 12661006). Tyrosine phosphorylated ROS1 is downregulated in response to crizotinib treatment (PMID : 24971615)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Phospho-ROS1 (Tyr2274/2334) antibody [EPR28163-133] (<a href='/en-us/products/primary-antibodies/phospho-ros1-tyr2274-2334-antibody-epr28163-133-ab324617'>ab324617</a>) at 1/1000 dilution

Lane 1:

Untreated U-118 MG (human brain glioblastoma cell) whole cell lysate (untreated membrane) at 50 µg

Lane 2:

U-118 MG treated with 250 nM crizotinib for 4 hours whole cell lysate (untreated membrane) at 50 µg

Lane 3:

Untreated U-118 MG whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 4:

U-118 MG treated with 250 nM crizotinib for 4 hours whole cell lysate (alkaline phosphatase treated membrane) at 50 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: 110 kDa,36 kDa

false

Exposure time: 180s