PA5-17648

antibody from Invitrogen Antibodies

Targeting: TP53 LFS1, p53

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-17648

Provider

Invitrogen Antibodies

Product name

Acetyl-p53 (Lys382) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody. This antibody is not cross-reactive with other acetylated proteins.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Acetyl-p53 (Lys382) Polyclonal Antibody (Product # PA5-17648) and a 53 and 44 kDa band corresponding to Cellular tumor antigen p53 (Lys382) was observed in MCF7 treated with 0.5 µM of Doxorubicin for 24 hours which further increased upon treatment with 400 nM of Trichostatin A along with 0.5 µM of Doxorubicin for 24 hours and absent in untreated MCF7, K-562 and SK-OV-3 along with an uncharacterized band at 35 kDa(*). Nuclear enriched extracts (30 µg lysate) of MCF7 (Lane 1), MCF7 treated with 0.5 µM of Doxorubicin for 24 hours (Lane 2), MCF7 (Lane 3), MCF7 treated with 0.5 µM of Doxorubicin and 400 nM of Trichostatin A for 24 hours(Lane 4), K-562 (Lane 5) and SK-O-V3 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescentdetection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Cellular tumor antigen p53 was performed using 70% confluent log phase MCF7 cells treated with 0.5 µM Doxorubicin and 400 nM Trichostatin A for 24 hours. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Acetyl-p53 (Lys382) Polyclonal Antibody (Product # PA5-17648) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing nuclear localization. Panel e represents untreated MCF7 cells showing no expression of Acetyl-p53. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.