Western blot abreview for Anti-p53 antibody [Y5]

Inconclusive
Abreviews
Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing (13 %)
Sample
Human Cell lysate - whole cell (Human HCT116 and MEF cell lysates)
Specification
Human HCT116 and MEF cell lysates
Treatment
DMSO control and 7.5 ´M CPT-11 treatment for 24 hrs
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Other product details

Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 2% Milk in PBS-T buffer
Dilution
1/1000

Secondary antibody

Name
Dilution
1/10000

Detection

Detection method
ECL
Bands
Specific: 55 kDa Non-specific: 130, 40, 35, 16 kDa
Exposure
30 second(s)
Negative control
HCT116 p53-/- lysates
Positive control
HCT116 p53wt treated with CPT-11 (7.5 ´M for 24 hrs) lysates

Additional data

Additional Notes
Specific signal at 55 kDa only after CPT-11 treatment, which induces p53 expression. Intense unspecific signal in both HCT116 p53wt and p53-/- cells at 35 and 40 kDa. Specificity of ab32049 is reduced compared to a non Abcam anti-p53 antibody. ab32049 additionally does not detect murine p53.
Abcam response
Thank you for your valuable feedback. ab32049 has been used to detect p53 in Human lung adenocarcinoma (A549) cells by Western blot. Your data suggests that ab32049 may not be suitable to detect endogenous p53 - without induction - by WB in Human cell lines under these experimental conditions. We hope this information is useful to our customers and we welcome further feedback.

Mr. Christian Marx

Verified customer

Submitted Jan 16 2014

Sign up