Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PRC1 with Purified 51248 at 1:30 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PRC1 with Purified 238427 at 1:50 dilution (5.06 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

All lanes : Anti-PRC1 antibody [EP1513Y] (ab51248) at 1/1000 dilution (Purified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 3 : C2C12 (Mouse myoblasts myoblast) whole cell lysate
Lane 4 : Mouse kidney lysate
Lane 5 : Rat kidney lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 72 kDa

ab51248 recognise two isoforms of PRC1. But we are unsure how to define the extra bands at ~50kDa.

ab51248 staining PRC1 in human breast cancer cells by ICC/IF. The cells were paraformaldehyde fixed and blocked in 1% serum for 1 hour at 37°C without permeation step. The primary antibody was diluted 1/100 (PBS) and incubated with sample for 1 hour at 20°C. An Alexa Fluor® 488 conjugated donkey polyclonal to rabbit IgG, diluted 1/200 was used as secondary.

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