产品名称Anti-p53抗体[DO-1] - ChIP Grade
参阅全部 p53 一抗
描述小鼠单克隆抗体[DO-1] to p53 - ChIP Grade
经测试应用适用于: ChIP, ICC/IF, ELISA, IHC-P, IHC-Fr, IP, WB, Flow Cytmore details
不与反应: Mouse, Rat
Recombinant full length protein corresponding to Human p53 (N terminal). The epitope is within aa 20-25.
Database link: P04637
- WB: A431, MCF7, HEK-293, DU145, MDA-MB-361 cell lysate. IHC-P: Human colon adenocarcinoma FFPE tissue sections. ICC/IF: A431 cells. ChIP: HEK-293 cell chromatin.
This monoclonal p53 antibody has been knockout validated in Western blot. The expected band for p53 was observed in wild type HAP1 cell lysate and the band was not seen in TP53 knockout HAP1 cell lysate.
This antibody clone is manufactured by Abcam.
存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Preservative: 0.02% Sodium azide
Some batches contain 6.97% L-Arginine as a stabilizing agent. For lot-specific buffer information, please contact our Scientific Support team.
Concentration information loading...
纯度Protein G purified
ChIP Related Products
Our Abpromise guarantee covers the use of ab1101 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent concentration.
Use at an assay dependent concentration.
|ICC/IF||Use at an assay dependent concentration.|
|ELISA||Use a concentration of 1 - 5 µg/ml.|
|IHC-P||Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.|
|IP||Use a concentration of 1 - 5 µg/ml.|
|WB||1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).|
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
功能Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. Involved in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for this process. One of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression. Implicated in Notch signaling cross-over. Isoform 2 enhances the transactivation activity of isoform 1 from some but not all TP53-inducible promoters. Isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis.
组织特异性Ubiquitous. Isoforms are expressed in a wide range of normal tissues but in a tissue-dependent manner. Isoform 2 is expressed in most normal tissues but is not detected in brain, lung, prostate, muscle, fetal brain, spinal cord and fetal liver. Isoform 3 is expressed in most normal tissues but is not detected in lung, spleen, testis, fetal brain, spinal cord and fetal liver. Isoform 7 is expressed in most normal tissues but is not detected in prostate, uterus, skeletal muscle and breast. Isoform 8 is detected only in colon, bone marrow, testis, fetal brain and intestine. Isoform 9 is expressed in most normal tissues but is not detected in brain, heart, lung, fetal liver, salivary gland, breast or intestine.
疾病相关Note=TP53 is found in increased amounts in a wide variety of transformed cells. TP53 is frequently mutated or inactivated in about 60% of cancers. TP53 defects are found in Barrett metaplasia a condition in which the normally stratified squamous epithelium of the lower esophagus is replaced by a metaplastic columnar epithelium. The condition develops as a complication in approximately 10% of patients with chronic gastroesophageal reflux disease and predisposes to the development of esophageal adenocarcinoma.
Defects in TP53 are a cause of esophageal cancer (ESCR) [MIM:133239].
Defects in TP53 are a cause of Li-Fraumeni syndrome (LFS) [MIM:151623]. LFS is an autosomal dominant familial cancer syndrome that in its classic form is defined by the existence of a proband affected by a sarcoma before 45 years with a first degree relative affected by any tumor before 45 years and another first degree relative with any tumor before 45 years or a sarcoma at any age. Other clinical definitions for LFS have been proposed (PubMed:8118819 and PubMed:8718514) and called Li-Fraumeni like syndrome (LFL). In these families affected relatives develop a diverse set of malignancies at unusually early ages. Four types of cancers account for 80% of tumors occurring in TP53 germline mutation carriers: breast cancers, soft tissue and bone sarcomas, brain tumors (astrocytomas) and adrenocortical carcinomas. Less frequent tumors include choroid plexus carcinoma or papilloma before the age of 15, rhabdomyosarcoma before the age of 5, leukemia, Wilms tumor, malignant phyllodes tumor, colorectal and gastric cancers.
Defects in TP53 are involved in head and neck squamous cell carcinomas (HNSCC) [MIM:275355]; also known as squamous cell carcinoma of the head and neck.
Defects in TP53 are a cause of lung cancer (LNCR) [MIM:211980].
Defects in TP53 are a cause of choroid plexus papilloma (CPLPA) [MIM:260500]. Choroid plexus papilloma is a slow-growing benign tumor of the choroid plexus that often invades the leptomeninges. In children it is usually in a lateral ventricle but in adults it is more often in the fourth ventricle. Hydrocephalus is common, either from obstruction or from tumor secretion of cerebrospinal fluid. If it undergoes malignant transformation it is called a choroid plexus carcinoma. Primary choroid plexus tumors are rare and usually occur in early childhood.
Defects in TP53 are a cause of adrenocortical carcinoma (ADCC) [MIM:202300]. ADCC is a rare childhood tumor of the adrenal cortex. It occurs with increased frequency in patients with the Beckwith-Wiedemann syndrome and is a component tumor in Li-Fraumeni syndrome.
序列相似性Belongs to the p53 family.
结构域The nuclear export signal acts as a transcriptional repression domain. The TADI and TADII motifs (residues 17 to 25 and 48 to 56) correspond both to 9aaTAD motifs which are transactivation domains present in a large number of yeast and animal transcription factors.
翻译后修饰Acetylated. Acetylation of Lys-382 by CREBBP enhances transcriptional activity. Deacetylation of Lys-382 by SIRT1 impairs its ability to induce proapoptotic program and modulate cell senescence.
Phosphorylation on Ser residues mediates transcriptional activation. Phosphorylated by HIPK1 (By similarity). Phosphorylation at Ser-9 by HIPK4 increases repression activity on BIRC5 promoter. Phosphorylated on Thr-18 by VRK1. Phosphorylated on Ser-20 by CHEK2 in response to DNA damage, which prevents ubiquitination by MDM2. Phosphorylated on Thr-55 by TAF1, which promotes MDM2-mediated degradation. Phosphorylated on Ser-46 by HIPK2 upon UV irradiation. Phosphorylation on Ser-46 is required for acetylation by CREBBP. Phosphorylated on Ser-392 following UV but not gamma irradiation. Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylated on Ser-15 upon ultraviolet irradiation; which is enhanced by interaction with BANP.
Dephosphorylated by PP2A-PPP2R5C holoenzyme at Thr-55. SV40 small T antigen inhibits the dephosphorylation by the AC form of PP2A.
May be O-glycosylated in the C-terminal basic region. Studied in EB-1 cell line.
Ubiquitinated by MDM2 and SYVN1, which leads to proteasomal degradation. Ubiquitinated by RFWD3, which works in cooperation with MDM2 and may catalyze the formation of short polyubiquitin chains on p53/TP53 that are not targeted to the proteasome. Ubiquitinated by MKRN1 at Lys-291 and Lys-292, which leads to proteasomal degradation. Deubiquitinated by USP10, leading to its stabilization. Ubiquitinated by TRIM24, which leads to proteasomal degradation. Ubiquitination by TOPORS induces degradation. Deubiquitination by USP7, leading to stabilization. Isoform 4 is monoubiquitinated in an MDM2-independent manner.
Monomethylated at Lys-372 by SETD7, leading to stabilization and increased transcriptional activation. Monomethylated at Lys-370 by SMYD2, leading to decreased DNA-binding activity and subsequent transcriptional regulation activity. Lys-372 monomethylation prevents interaction with SMYD2 and subsequent monomethylation at Lys-370. Dimethylated at Lys-373 by EHMT1 and EHMT2. Monomethylated at Lys-382 by SETD8, promoting interaction with L3MBTL1 and leading to repress transcriptional activity. Demethylation of dimethylated Lys-370 by KDM1A prevents interaction with TP53BP1 and represses TP53-mediated transcriptional activation.
Sumoylated by SUMO1.
细胞定位Cytoplasm; Cytoplasm. Nucleus. Nucleus > PML body. Endoplasmic reticulum. Interaction with BANP promotes nuclear localization. Recruited into PML bodies together with CHEK2; Nucleus. Cytoplasm. Localized in both nucleus and cytoplasm in most cells. In some cells, forms foci in the nucleus that are different from nucleoli; Nucleus. Cytoplasm. Localized in the nucleus in most cells but found in the cytoplasm in some cells; Nucleus. Cytoplasm. Localized mainly in the nucleus with minor staining in the cytoplasm; Nucleus. Cytoplasm. Predominantly nuclear but localizes to the cytoplasm when expressed with isoform 4 and Nucleus. Cytoplasm. Predominantly nuclear but translocates to the cytoplasm following cell stress.
- Information by UniProt
- Antigen NY-CO-13 antibody
- BCC7 antibody
- Cellular tumor antigen p53 antibody
IHC image of ab1101 staining p53 in human colon adenocarcinoma formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab1101, 1/500 dilution, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. High magnification of the tumor region - T (lower right panel) and adjacent normal crypts - N (lower left panel) are shown.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
All lanes : Anti-p53 antibody [DO-1] - ChIP Grade (ab1101) at 0.4 µg/ml
Lane 1 : Extract of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells incubated with vehicle at 20 µg
Lane 2 : Extract of HEK-293T cells incubated with etoposide at 20 µg
Lane 3 : Extract of HEK-293T cells incubated with etoposide at 7.5 µg
Lane 4 : Lambda phosphatase (diluted 1/400)-treated extract of HEK-293T cells incubated with etoposide at 7.5 µg
Lane 5 : Lambda phosphatase (diluted 1/100)-treated extract of HEK-293T cells incubated with etoposide at 7.5 µg
Lane 6 : Lambda phosphatase (diluted 1/25)-treated extract of HEK-293T cells incubated with etoposide at 7.5 µg
Lane 7 : Extract of MCF7 (human breast adenocarcinoma cell line) cells incubated with vehicle at 20 µg
Lane 8 : Extract of MCF7 cells incubated 6 hours with camptothecin at 20 µg
Lane 9 : Extract of MCF7 cells incubated 16 hours with camptothecin at 20 µg
Lane 10 : Extract of MCF7 cells incubated 24 hours with camptothecin at 20 µg
All lanes : Goat anti mouse IgG(H&L)-HRP at 1/10000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Developed using the ECL technique with 30 second exposure.
Blocking: 5% milk in PBS for 3 hours at RT.
Primary antibody in 5% BSA + PBS was incubated overnight at 4°C.
Secondary antibody in 5% milk + PBS was incubated for 2 hours at RT.
ab1101 stained in A431 cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab1101 at 1 µg/ml and ab6046 (Rabbit polyclonal to beta Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150080 (pseudo-colored red) and ab150117 (colored green) used at 1 µg/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1 hour at room temperature
Chromatin was prepared from HEK-293 (human epithelial cell line from embryonic kidney) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 2 µg of ab1101, and 10 ml of protein A sepharose beads,10 ml of protein G sepharose beads. No IgG was added to the beads control. The immunoprecipitated DNA was quantified by real time PCR. Primers were as follows:
Bax-1, forward: GGGTTATCTCTTGGGCTCACAA.
Bax-1, reverse: GAGCTCTCCCCAGCGCA.
Bax-2, forward: TGG AGC TGC AGA GGA TGA TTG
Bax-2, reverse: CCA GTT GAA GTT GCC GTC AGA
PUMA, forward: ATG CCT GCC TCA CCT TCA TC
PUMA, reverse: TCA CAC GTC GCT CTC TCT AAA CC
p21-1, forward: GCT GTG GCT CTG ATT GGC TTT
p21-1, reverse: ACA GGC AGC CCA AGG ACA AA
p21-2, forward: CAT CCC CAC AGC AGA GGA GAA
p21-2, reverse: ACC CAG GCT TGG AGC AGC TA
p21-3, forward: GAG TCC TGT TTG CTT CTG GGC A
p21-3, reverse: CTG CAT TGG GGC TGC CTA TGT A
PCNA, forward: CCA CCA TAA AGC TGG GGC TT
PCNA, reverse: TCT CCC CGC CTC TTT GAC TC
Lanes 1, 5 and 9: Wild type HAP1 cell lysate (20 µg)
Lanes 2, 6 and 10: p53 knockout HAP1 cell lysate (20 µg)
Lanes 3, 7 and 11: A431 cell lysate (20 µg)
Lanes 4, 8 and 12: Saos-2 cell lysate (20 µg)
Lanes 1, 2, 3 and 4: Green signal from target – ab1101 observed at 53 kDa
Lanes 5, 6, 7 and 8: Red signal from loading control – ab181602 observed at 37 kDa
Lanes 9, 10, 11 and 12: Merged (red and green) signal
ab1101 was shown to specifically react with p53 in wild type HAP1 cells No band was observed in p53 knockout HAP1 cell lysates. Wild type and p53 knockout samples were subjected to SDS-PAGE. ab1101 and ab181602 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
All lanes : Anti-p53 antibody [DO-1] - ChIP Grade (ab1101) at 1/1000 dilution
Lane 1 : HEK-293 (human embryonic kidney cell line) whole cell lysate
Lane 2 : DU 145 (human prostate carcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Predicted band size: 53 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with ab1101 overnight at 4°C. Antibody binding was detected using ab175783 at a 1/10,000 dilution for 1 hour at room temperature and then imaged using the Licor Odyssey CLx.
All lanes : Anti-p53 antibody [DO-1] - ChIP Grade (ab1101) at 2.5 µg/ml
Lane 1 : MDA-MB-361 (human breast adenocarcinoma cell line) whole cell lysate
Lane 2 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
Lane 3 : A549 (human lung adenocarcinoma epithelial cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Additional bands at: 55 kDa (possible post-translational modification)
Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab1101 (red line). The cells were fixed with 4% paraformaldehyde for 10 minutes and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab1101, 1/50 dilution) for 30 minutes at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) ab96879 at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] ab91361, 1 µg/1 x 106 cells, used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.
This product has been referenced in:
- Wang L et al. MicroRNA-188-3p is involved in sevoflurane anesthesia-induced neuroapoptosis by targeting MDM2. Mol Med Rep 17:4229-4236 (2018). Read more (PubMed: 29344658) »
- Waldherr M et al. Use of HuH6 and other human-derived hepatoma lines for the detection of genotoxins: a new hope for laboratory animals? Arch Toxicol 92:921-934 (2018). Read more (PubMed: 29218508) »