TP53 Antibody (AMM00025)

Catalog#: AMM00025

• Size: 100ug

Human Daudi

Human cell lines

Human U2OS, PLKO

• Description of Target: Tumor protein p53 responds to diverse cellular stresses to regulate target genes that induce cell cycle arrest, apoptosis, senescence, DNA repair, or changes in metabolism. p53 protein is expressed at low level in normal cells and at a high level in a variety of transformed cell lines, where it's believed to contribute to transformation and malignancy. p53 is a DNA-binding protein containing transcription activation, DNA-binding, and oligomerization domains. It is postulated to bind to a p53-binding site and activate expression of downstream genes that inhibit growth and/or invasion, and thus function as a tumor suppressor. Mutants of p53 that frequently occur in a number of different human cancers fail to bind the consensus DNA binding site, and hence cause the loss of tumor suppressor activity. Alterations of this gene occur not only as somatic mutations in human malignancies, but also as germline mutations in some cancer-prone families with Li-Fraumeni syndrome
• Gene Symbol: TP53
• Alias Symbols: P53; LFS1; TRP53; FLJ92943; TP53
• Protein Accession#: NP_000537
• Nucleotide Accession#: NM_000546
• Swissprot Id: P04637
• Host: Mouse
• Protein Size (# AA): 393
• Molecular Weight: 53kDa
• Application: WB, ChIP
• Product Format: Lyophilized powder
• Isotype: Mouse IgG
• Reconstitution and Storage: Add 100ul of distilled water. Final anti-TP53 antibody concentration is 1 mg/ml in PBS buffer. For longer periods of storage, store at -20°C. Avoid repeat freeze-thaw cycles.
• Key Reference: Khanna,K.K., et al., Nat. Genet. 20 (4), 398-400 (1998)

Customer Reviews for TP53 Antibody (AMM00025) tested with U20S, PLKO cells in Chromatin Immunoprecipitation

CAT# AMM00025
-submitted by Nick Barlev
University of Leicester

U20S (p53+) cells were treated with 0.5 uM Doxorubicin for 14 hrs to induce DNA damage and hence activate p53. In parallel, PLKO cells (U2OS cells with stable shRNA-mediated knockdown of p53) were treated similarly and were used as negative control. Thedata for p21 promoter were normalised to actin (control for non-specific binding of DNA to the antibodies).

Product Review: TP53 Antibody (AMM00025) in PLKO AND U2OS cells using ChIp Assays

Product Page Link: TP53 Antibody (AMM00025)

Data provided by: Dr. Barlev, University of Leicester

Figure 1. Binding of p53-specific antibodies to the p21 promoter. U20S (p53+) cells were treated with 0.5 uM Doxorubicin for 14 hrs to induce DNA damage and hence activate p53. In parallel, PLKO cells (U2OS cells with stable shRNA-mediated knockdown of p53) were treated similarly and were used as negative control. Thedata for p21 promoter were normalised to actin (control for non-specific binding of DNA to the antibodies).

Product Review: TP53 Antibody (AMM00025) in MIA PaCa-2 human pancreatic cancer cell line, MDA-MB-231 Human breast carcinoma cell line and HUH 7 hepatocarcinoma cell line using Western blot

Product page for TP53 Antibody (AMM00025)

Researcher:Andrei L. Gartel, University of Illinois at Chicago
Application: Western blotting
Species+tissue/cell type: Lane 1: 25ug MIA PaCa-2 cell lysate Lane 2: 25ug MDA-MB-231 cell lysate Lane 3: 25ug Huh-7 cell lysate
Primary antibody dilution: 1:2000
Secondary antibody: Anti-rabbit-HRP
Secondary antibody dilution: 1:5000

Questionnaire:
How do Aviva’s reagents play a role in your experimental goals?	We haven't used them until the trial.
How would you rate this antibody on a scale from 1-5 (5=best) and why?	4: -some background bands
Would you use this antibody in future experiments?	May be
Have you used another antibody which has worked in your application?	Yes
Do you believe the information about the reagent on Aviva’s website is correct?	Yes
If the antibody works, do you plan to use it in future experiments or to publish your data? Why or why not?	May be.
How did you store the antibody after re-suspension?	4 degree C
Sample Description (please include 1) species type, and 2) cell/tissue type, and 3) how much protein loaded for each lane of your gel):	Human cancer cells; 25 ng
How many different experimental trials were conducted using the antibody sample?	One
How was this sample prepared?	Cells were lysed in 20mM Hepes, 1% Triton-X-100, 150mM NaCl, 1mM EDTA, 1mM EGTA, 100 mM NaF, 10mM Na4P2O7, 1mM Na3VO4, 0.2mM PMSF containing buffer.
Primary antibody dilution and incubation time:	1:2000; overnight at 4 degree C
Secondary antibody used and dilution and incubation time:	1:5000; 1 hr at room temp.; Jackson Immunoresearch
What controls were used in your experiment (positive/negative)?	Our own corresponding antibodies.
Please include your detailed WB Procedure/Protocol here:	- Samples were mix on 10% gel in 1X running buffer (Tris-Glycine-SDS). - Transfered to PVDF membrane in trannsfer buffer (Tris-Glycine-MeOH) for 24hrs at 48V. - Blocked in 5% milk (in TBST) for 1 hr at room temp. - Primary1:2000 overnight at 4 degree C. - Washed 3X 15 min in TBST. - Secondary 1:5000 1 hr at room temp. - Washed 3x 15 min in TBST. - Substrate (Thermo scientific #34080) for 5 min and expose film for 30 sec, 3 min or overnight.