Anti-p53 antibody [pAb122]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [pAb122] (AB90363)

ab90363 staining p53 in wild-type Hap1 cells (top panel) and p53 knockout Hap1 cells (bottom panel). The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% 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 with ab90363 at 1/500 dilution and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).

• WB

AbReview56174****

Western blot - Anti-p53 antibody [pAb122] (AB90363)

Blocked with 5% milk for 1 hour at 21^oC.

All lanes:

Western blot - Anti-p53 antibody [pAb122] (ab90363) at 1/2000 dilution

Lane 1:

HCT-116 cell lysate, untreated at 30 µg

Lane 2:

HCT-116 cell lysate + 10 µM CPT-11 for 24 hrs at 30 µg

Lane 3:

Negative control: HCT-116 cell lysate, untreated at 30 µg

Lane 4:

Negative control: HCT-116 cell lysate + 10 µM CPT-11 for 24 hrs at 30 µg

Secondary

All lanes:

HRP-conjugated rabbit anti-mouse polyclonal at 1/5000 dilution

Predicted band size: 43 kDa

Observed band size: 130 kDa,35 kDa,55 kDa

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Exposure time: 5min

This image is courtesy of an Abreview by Anke Rauch.

• WB

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Western blot - Anti-p53 antibody [pAb122] (AB90363)

All lanes:

Western blot - Anti-p53 antibody [pAb122] (ab90363) at 1/250 dilution

Lane 1:

A431

Lane 2:

Rat brain tissue extract

Lane 3:

Mouse brain tissue extract

Predicted band size: 43 kDa

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