Purified anti-p53 Antibody

Pricing & Availability
Clone
DO-7 (See other available formats)
Regulatory Status
RUO
Other Names
Tumor protein p53 (TP53)
Isotype
Mouse IgG2b, κ
1_DO-7_PURE_p53_Antibody_1_031817
Total cell lysate from Raw264.7 (lane 1) and MCF-7 (lane 2) (15 µg/lane) were resolved by electrophoresis (4-20% Tris-Glycine), transferred to nitrocellulose, and probed with 0.5 µg/mL purified anti-p53 antibody (clone DO-7). Proteins were visualized using an HRP goat anti-mouse-IgG secondary antibody (clone Poly4053) and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin antibody (clone 2F1-1) was used as a loading control. Lane M is the MW ladder.
  • 1_DO-7_PURE_p53_Antibody_1_031817
    Total cell lysate from Raw264.7 (lane 1) and MCF-7 (lane 2) (15 µg/lane) were resolved by electrophoresis (4-20% Tris-Glycine), transferred to nitrocellulose, and probed with 0.5 µg/mL purified anti-p53 antibody (clone DO-7). Proteins were visualized using an HRP goat anti-mouse-IgG secondary antibody (clone Poly4053) and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin antibody (clone 2F1-1) was used as a loading control. Lane M is the MW ladder.
  • 2_DO-7_PURE_p53_Antibody_4_031519
    Whole cell extracts (15 µg total protein) from HEK293, A431, and THP-1 were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to PVDF, and probed with 1.0 µg/mL (1:500 dilution) of Purified anti-p53 Antibody, clone DO-7, overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG Antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular Weight marker. Cell lysates were loaded in descending of predicted p53 expression; Predicted expression data was obtained from Human Protein Atlas.
  • 3_DO-7_PURE_p53_Antibody_2_031817
    BT474 cells were stained with anti-p53 (clone DO-7), followed by Alexa Fluor® 546 secondary antibody and DAPI (nuclei). Images were aquired on a Nikon FC300 inverted microscope at 20X magnification. Data provided by Dr. John Nolan, La Jolla Bioengineering Institute.
  • 4_DO-7_PURE_p53_Antibody_3_031817
    Immunofluorescence of Daudi cells with (A) Mouse IgG2b, κ Isotype control (Cat. No. 400302) or (B-D) TP-53 primary antibody. Alexa Fluor® 594 (Red) Goat anti-Mouse IgG (Cat. No. 405326) was used as secondary antibody. Nuclei were counterstained with DAPI (Blue, Cat. No. 422801). The image was captured with a 60X objective. Exposure time (Seconds) is 1/12. Concentrations for (A, B) is 5 µg/ml, (C) is 2 µg/ml, (D) is 1 µg/ml.
  • 5_DO-7_PURE_p53_Antibody_5_040320
    Chromatin Immunoprecipitations (ChIP) were done with fixed and sonicated chromatin samples from 293T cells treated with tunicamycin (2 µg/mL, 8 hr). ChIP was performed with 5 µg of chromatin and either 1 µg of purified anti-p53 antibody (clone DO-7, Cat. No. 645802), or equal amount of purified mouse IgG2b, κ isotype ctrl antibody (clone MPC-11, Cat. No. 400302). The enriched DNA was purified and quantified by real-time qPCR using SYBR Green and primers for the human CDKN1A promoter, and for a gene desert region on chromosome 12. The amount of immunoprecipitated DNA in each sample is represented as a percentage of the total amount of input chromatin, which is equivalent to 100%.
  • 6-DO-7_PURE_p53_Antibody_6_040323
    THP-1 cells (negative control, open histogram), or Daudi cells (positive control, filled histogram) were fixed and permeabilized using True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) and intracellularly stained with purified anti-p53 (clone DO-7), or purified mouse IgG2b, κ isotype control (open histogram, dashed line) (representative histogram for both cell lines) (Cat. No. 400302), followed by PE goat anti-mouse IgG (Cat. No. 405307).
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645801 25 µg $106.00
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645802 100 µg $226.00
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Description

p53 is a 53 kD protein that forms tetramers and functions as a tumor suppressor and transcriptional activator of genes that inhibit growth and/or invasion, cell cycle checkpoint after irradiation, DNA repair, apoptotic induction, signal transduction, and cell adhesion. This protein is localized to the nucleus when activated and can be upregulated by genotoxic or other cellular stresses. p53 is modified by phosphorylation, acetylation, ribosylation, ubiquitination, and sumoylation; ubiquination targets p53 for degradation via mdm2. This protein interacts with a variety of proteins including mdm2, mdmx, topoisomerase I, PML3, Bcl-XL, Bcl-2, Chk1, JNK, p38, HIPK2, CK2, DNA-PK, p300/CBP, PCAF, PARP1, and HDAC1-3. Mutant p53 associates with p63 and p73.

Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
p53 N-terminal linear epitope aa 20-25
Formulation
This antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
This antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, ChIP, ICFC - Verified
ELISA - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. Western blotting, suggested working dilution(s): Use 0.5 - 1.0 µg per mL antibody for each mini-gel. For immunocytochemistry, a concentration range of 2.0 - 5.0 μg/mL (Dilution 1:100 - 1:250) is recommended. The recommended starting antibody to chromatin ratio is 1 µg of antibody and 5 µg of chromatin per IP. Optimal antibody to chromatin ratio should be determined by the end user. For intracellular flow cytometric staining, the suggested use of this reagent is ≤ 0.03 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application References

(PubMed link indicates BioLegend citation)
  1. Vojtesek B, et al. 1992. J. Immunol. Methods 151:237.
  2. Stephen CW, et al. 1995. J. Mol. Biol. 248:58.
Product Citations
  1. Senosain MF, et al. 2021. Sci Rep. 11:14424. PubMed
RRID
AB_2206461 (BioLegend Cat. No. 645801)
AB_2206461 (BioLegend Cat. No. 645802)

Antigen Details

Structure
Tetramer; 53 kD
Distribution

Nuclear when activated

Function
Tumor suppressor, transcription factor regulates cell cycle checkpoint, apoptosis, DNA integrity
Interaction
mdm2, mdmx, TopoI, PML3, Bcl-XL, Bcl-2, Chk1, JNK, p38, HIPK2, CK2, DNA-PK, p300/CBP, PCAF, PARP1, HDAC1-3; mutant p53 associates with p63, p73
Cell Type
Mesenchymal Stem Cells
Biology Area
Cell Biology, DNA Repair/Replication, Stem Cells
Gene ID
7157 View all products for this Gene ID
UniProt
View information about p53 on UniProt.org

Other Formats

View All p53 Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-p53 DO-7 WB,ICC,ChIP,ICFC,ELISA
FITC anti-p53 DO-7 ICFC
PE anti-p53 DO-7 ICFC
Go To Top Version: 7    Revision Date: 04/03/2023

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