xCT / SLC7A11 Recombinant Mouse Monoclonal Antibody [A7C6-R]

Western blot analysis of xCT / SLC7A11 on HCT116 cell lysates with Mouse anti-xCT / SLC7A11 antibody (HA601071) at 1/5,000 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 1 minute;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601071) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

Western blot analysis of xCT / SLC7A11 on NIH/3T3 cell lysates with Mouse anti-xCT / SLC7A11 antibody (HA601071) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 55 kDa
Observed band size: 55 kDa

Exposure time: 1 minute;

8% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601071) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Mouse anti-xCT / SLC7A11 antibody (HA601071) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA601071) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry analysis of HT-29 cells labeling xCT / SLC7A11 with Mouse anti-xCT / SLC7A11 antibody (HA601071) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-xCT / SLC7A11 antibody (HA601071) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

Flow cytometric analysis of HT-29 cells labeling xCT / SLC7A11.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA601071, 1/1,000) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for 30 minutes, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).