Anti-Cdc20 antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cdc20 antibody (AB26483)

ab26483 staining Cdc20 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab26483 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min).Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

Project1671****

Western blot - Anti-Cdc20 antibody (AB26483)

ab26483 consistently detects a band at 55-60 kDa but we also see bands of ~35 and 70 kDa that appear with varying intensity in some of our extracts. We cannot find an explanation for this and are unsure of the nature of these crossreacting bands.

All lanes:

Western blot - Anti-Cdc20 antibody (ab26483) at 1 µg/mL

Lane 1:

Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate (<a href='/en-us/products/unavailable/raji-human-burkitt-s-lymphoma-cell-line-whole-cell-lysate-ab7908'>ab7908</a>) at 20 µg

Lane 2:

Western blot - THP-1 whole cell lysate (<a href='/en-us/products/cell-lysates/thp-1-whole-cell-lysate-ab7913'>ab7913</a>) at 20 µg

Lane 3:

Western blot - HEK-293 whole cell lysate (<a href='/en-us/products/cell-lysates/hek-293-whole-cell-lysate-ab7902'>ab7902</a>) at 20 µg

Lane 4:

Western blot - HL-60 whole cell lysate (<a href='/en-us/products/cell-lysates/hl-60-whole-cell-lysate-ab7914'>ab7914</a>) at 20 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilution

Predicted band size: 55 kDa

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