KRAS Mouse Monoclonal Antibody [A8E5]

Western blot analysis of KRAS on different lysates with Mouse anti-KRAS antibody (HA601059) at 1/2,000 dilution.

Lane 1: HCT 116 cell lysate
Lane 2: HEK-293 cell lysate
Lane 3: 22RV1 cell lysate
Lane 4: Mouse brain tissue lysate
Lane 5: Mouse kidney tissue lysate
Lane 6: Rat brain tissue lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 22 kDa
Observed band size: 22 kDa

Exposure time: 3 minutes; ECL: K1801;

4-20% 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 (HA601059) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of Jurkat cells labeling KRAS with Mouse anti-KRAS antibody (HA601059) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-KRAS antibody (HA601059) 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) were used as the secondary antibody at 1/1,000 dilution.

Immunocytochemistry analysis of NIH/3T3 cells labeling KRAS with Mouse anti-KRAS antibody (HA601059) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-KRAS antibody (HA601059) at 1/100 dilution in 2% BSA 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.

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Mouse anti-KRAS antibody (HA601059) at 1/1,100 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 (HA601059) at 1/1,100 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.

Flow cytometric analysis of Jurkat cells labeling KRAS.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA601059, 1/1,000) (red) compared with Mouse IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, 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).

Flow cytometric analysis of NIH/3T3 cells labeling KRAS.

Cells were fixed and permeabilized, and then blocked with 2% negative goat serum for 15 minutes at room temperature.Then stained with the primary antibody (HA601059, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, 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).