All lanes : Anti-MCM6 antibody - C-terminal (ab236449) at 1/2000 dilution

Lane 1 : U-2 OS (human bone osteosarcoma epithelial cell line) siCtrl
Lane 2 : U-2 OS (human bone osteosarcoma epithelial cell line) siMCM6
Lane 3 : MEF (mouse embryonic fibroblast cell line) cell lysate
Lane 4 : INS-1 (rat cell line) cell lysate
Lane 5 : DT40 (chicken lymphoma cell line) cell lysate
Lane 6 : Xenopus laevis lysate

Lysates/proteins at 40 µg per lane.

Predicted band size: 93 kDa

Block: TBS-T 5% milk. Incubated overnight with primary antibody.

Treated with control or MCM6 siRNA to show specificity.

Top panel: U-2 OS (Human bone osteosarcoma epithelial cell line) cells stained for MCM6 using ab236449 at a dilution of 1/2000 in ICC/IF.

Middle panel: DAPI.

Bottom panel: Merged image.

Cells were fixed using 2% PFA. Incubated with primary antibody overnight at +4°C.

Flow Cytometry analysis of U-2 OS (Human bone osteosarcoma epithelial cell line) cells, labeling MCM6 with ab236449 at a 1/200 dilution (green) compared to a negative control (no primary antibody) (blue).

Cells  were fixed in PFA 1% for 10 minutes, permeabilized with Triton X-100 and incubated with the primary antibody overnight.

MCM6 was immunoprecipitated from U-2 OS (Human bone osteosarcoma epithelial cell line) whole cell lysate using 3 µl of ab236449.

Lane 1: U-2 OS whole cell lysate (Input).

Lane 2: Control IgG in U-2 OS whole cell lysate.

Lane 3: ab236449 in U-2 OS whole cell lysate.

For western blotting ab236449 was used at a 1/2000 dilution.