Anti-xCT antibody [3A12] (BSA and Azide free)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-xCT antibody [3A12] (BSA and Azide free) (AB302919)

This data was developed using ab300667, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A549 (human lung carcinoma epithelial cell) cells labelling xCT with ab300667 at 1/50 (21.18 ug/ml) dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing membraneous and cytoplasmic staining in A549 cell line.Low expression : colo 205 (PMID : 31000598). is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 10 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor® 488) at 1/1000 2 ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-xCT antibody [3A12] (BSA and Azide free) (AB302919)

This data was developed using ab300667, the same antibody clone in a different buffer formulation.xCT was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug with ab300667 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300667 at 1/1000 dilution. Goat Anti-Rat IgG (H+L), HRP) (ab205720) was used at 1/10000 dilution. Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug Lane 2 : ab300667 IP in HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate Lane 3 : Rat IgG2a, kappa monoclonal- Isotype Control (ab18450) instead of ab300667 in HepG2 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 seconds

• WB

Supplier Data

Western blot - Anti-xCT antibody [3A12] (BSA and Azide free) (AB302919)

This data was developed using ab300667, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. Low expression : SW480 and colo 205 (PMID : 31000598). Samples are non-boiled as boiling may cause protein aggregation. The expression profile observed is consistent with what has been described in the literature ( PMID : 32188872). Exposure time : 26 seconds

All lanes:

Western blot - Anti-xCT antibody [3A12] (<a href='/en-us/products/primary-antibodies/xct-antibody-3a12-ab300667'>ab300667</a>) at 1/1000 dilution

Lane 1:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

SW480 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 5:

COLO 205 (human colon; derived from metastatic site: ascites epithelial), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/10000 dilution

Observed band size: 55 kDa,35 kDa

false

Exposure time: 26s