p53 Antibody Sampler Kit

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p53 Antibody Sampler Kit

品牌： CST

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产品介绍

产品信息

荧光素标记

Unconjugated

抗原名称

p53

简单描述

The p53 Antibody Sampler Kit provides an economical means of detecting p53 activity using modification-specific and control antibodies. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

研究领域

癌症,细胞生物学,神经科学

应用

目标/特异性

Specificity/Sensitivity

p53 (7F5) Rabbit mAb detects endogenous levels of total p53 protein. This antibody binding has been mapped to the amino terminus region of human p53 protein.Acetyl- and phospho-specific antibodies detect p53 only when phosphorylated or acetylated at the specified site.

背景

The p53 tumor suppressor protein plays a major role in cellular response to DNA damage and other genomic aberrations. Activation of p53 can lead to either cell cycle arrest and DNA repair or apoptosis (1). p53 is phosphorylated at multiple sites in vivo and by several different protein kinases in vitro (2,3). DNA damage induces phosphorylation of p53 at Ser15 and Ser20 and leads to a reduced interaction between p53 and its negative regulator, the oncoprotein MDM2 (4). MDM2 inhibits p53 accumulation by targeting it for ubiquitination and proteasomal degradation (5,6). p53 can be phosphorylated by ATM, ATR, and DNA-PK at Ser15 and Ser37. Phosphorylation impairs the ability of MDM2 to bind p53, promoting both the accumulation and activation of p53 in response to DNA damage (4,7). Chk2 and Chk1 can phosphorylate p53 at Ser20, enhancing its tetramerization, stability, and activity (8,9). p53 is phosphorylated at Ser392 in vivo (10,11) and by CAK in vitro (11). Phosphorylation of p53 at Ser392 is increased in human tumors (12) and has been reported to influence the growth suppressor function, DNA binding, and transcriptional activation of p53 (10,13,14). p53 is phosphorylated at Ser6 and Ser9 by CK1δ and CK1ε both in vitro and in vivo (13,15). Phosphorylation of p53 at Ser46 regulates the ability of p53 to induce apoptosis (16). Acetylation of p53 is mediated by p300 and CBP acetyltransferases. Inhibition of deacetylation suppressing MDM2 from recruiting HDAC1 complex by p19 (ARF) stabilizes p53. Acetylation appears to play a positive role in the accumulation of p53 protein in stress response (17). Following DNA damage, human p53 becomes acetylated at Lys382 (Lys379 in mouse) in vivo to enhance p53-DNA binding (18). Deacetylation of p53 occurs through interaction with the SIRT1 protein, a deacetylase that may be involved in cellular aging and the DNA damage response (19). 1.Levine, A.J. (1997) Cell 88, 323-31. 2.Meek, D.W. (1994) Semin Cancer Biol 5, 203-10. 3.Milczarek, G.J. et al. (1997) Life Sci 60, 1-11. 4.Shieh, S.Y. et al. (1997) Cell 91, 325-34. 5.Chehab, N.H. et al. (1999) Proc Natl Acad Sci U S A 96, 13777-82. 6.Honda, R. et al. (1997) FEBS Lett 420, 25-7. 7.Tibbetts, R.S. et al. (1999) Genes Dev 13, 152-7. 8.Shieh, S.Y. et al. (1999) EMBO J 18, 1815-23. 9.Hirao, A. et al. (2000) Science 287, 1824-7. 10.Hao, M. et al. (1996) J Biol Chem 271, 29380-5. 11.Lu, H. et al. (1997) Mol Cell Biol 17, 5923-34. 12.Ullrich, S.J. et al. (1993) Proc Natl Acad Sci U S A 90, 5954-8. 13.Kohn, K.W. (1999) Mol Biol Cell 10, 2703-34. 14.Lohrum, M. and Scheidtmann, K.H. (1996) Oncogene 13, 2527-39. 15.Knippschild, U. et al. (1997) Oncogene 15, 1727-36. 16.Oda, K. et al. (2000) Cell 102, 849-62. 17.Ito, A. et al. (2001) EMBO J 20, 1331-40. 18.Sakaguchi, K. et al. (1998) Genes Dev 12, 2831-41. 19.Solomon, J.M. et al. (2006) Mol Cell Biol 26, 28-38.

研究领域

癌症,细胞生物学,神经科学

翻译后修饰

phosphate

数据库链接

Entrez-Gene ID

7157

UniProt ID

P04637

参考图片

Immunoprecipitation of Phospho-p53 (Ser15) protein from HT-29 + Etoposide #2200 (25µM, Overnight) cell extracts. Lane 1 is 10% input, lane 2 is Normal Rabbit IgG #2729, and lane 3 is Phospho-p53 (Ser15) Antibody. Western blot analysis was performed using Phospho-p53 (Ser15) Antibody. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as a secondary antibody. Note that Phospho-p53 (Ser15) Antibody cross-reacts with some proteins present in the Blue Prestained Protein Marker, Broad Range (11-250 kDa) #59329. In this figure, only proteins present in the Biotinylated Protein Ladder Detection Pack #7727 are labeled.

Western blot analysis of extracts from MCF-7 cells treated with etoposide for the indicated times, using Phospho-p53 (Ser46) Antibody.

Western blot analysis of extracts from HeLa cells, untreated or treated with both trichostatin A #9950 (400 nM for 24 hours), and doxorubicin (0.5 µM for 24 hours) using Acetyl-p53 (Lys382) Antibody alone (A), antibody preincubated with a non-acetylated Lys382 peptide (B), or antibody preincubated with an acetylated Lys382 peptide (C).

Western blot analysis of extracts from HT29 cells treated with nocodazole (50 ng/ml, 24h), calyculin A #9902 (100 nM, 10 min), or lambda Phosphatase NEB#P0753 (10,000 units/ml, 1 h), using Phospho-p53 (Ser33) Antibody (upper) or p53 (1C12) Mouse mAb #2524 (lower).

Western blot analysis of extracts from 293 and COS cells, using p53 (7F5) Rabbit mAb.

Western blot analysis of extracts from NIH/3T3 (left) and MCF-7 (right) cells, untreated, doxorubicin-treated (0.5 μM, 24 hours) or doxorubicin and trichostatin A-treated (TSA, #9950, 400 nM, 24 hours), using Acetyl-p53 (Lys379) Antibody.

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Western blot analysis of increasing amounts of a p53 fusion protein, untreated or phosphorylated by CKII, using Phospho-p53 (Ser392) Antibody (upper) or p53 Antibody #9282 (lower).

Western blot analysis of a p53 fusion protein, untreated or phosphorylated by DNA-PK, using Phospho-p53 (Ser15) Antibody (upper) and p53 Antibody #9282 (lower).

Western blot analysis of extracts from COS cells treated with UV or MMS for the indicated times, using Phospho-p53 (Ser20) Antibody (upper) or p53 Antibody #9282 (lower).

Western blot analysis of extracts from COS cells treated with UV or MMS for the indicated times, using Phospho-p53 (Ser9) Antibody (upper) or p53 Antibody #9282 (lower).

Immunoprecipitation of extracts from MCF-7 cells treated with etoposide under nondenaturing conditions, using Phospho-p53 (Ser46) Antibody, followed by Western blot analysis using a monoclonal p53 antibody.

Western blot analysis of extracts from HeLa cells, untreated, trichostatin A-treated #9950 (400 nM for 24 hours), doxorubicin-treated (0.5 µM for 24 hours), or both, using Acetyl-p53 (Lys382) Antibody (top) or p53 Antibody #2524 (bottom).

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-p53 (Ser33) Antibody.

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using p53 (7F5) Rabbit mAb.

Western blot analysis of extracts from Mv1Lu cells, untreated or hydroxyurea-treated (20 mM), using Phospho-p53 (Ser392) Antibody.

Western blot analysis of a p53 fusion protein, untreated or phosphorylated by CK1 or CK2, using Phospho-p53 (Ser9) Antibody (upper) or p53 Antibody #9282 (lower).

Confocal immunofluorescent analysis of MCF-7 cells, untreated (left) or etoposide-treated (right), using Phospho-p53 (Ser46) Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red).

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing nuclear localization, using Phospho-p53 (Ser33) Antibody.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using p53 (7F5) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human lung dysplasia of alveolar cells, using Phospho-p53 (Ser33) Antibody in the presence of control peptide (left) or antigen specific peptide (right),

Immunohistochemical analysis of paraffin-embedded HT-29 (left) and SaOs-2 (right) cells, using p53 (7F5) Rabbit mAb. Note the lack of staining in p53-negative SaOs-2 cells.

Confocal Immunofluorescent analysis of HT-29 cells using p53 (7F5) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

Flow cytometric analysis of HT-29 cells using p53 (7F5) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells treated with UV (100 J/m2 followed by a 3 hour recovery) and either p53 (7F5) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CDKN1A Promoter Primers #6449, human MDM2 intron 2 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Simple Western™ analysis of lysates (1 mg/mL) from COS-7 cells using p53 (7F5) Rabbit mAb #2527. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Western blot analysis of extracts from PC12 cells treated with UV for the indicated times, using Phospho-p53 (Ser15) Antibody.

Flow cytometric analysis of HT-29 cells, untreated (blue) or UV-treated (green), using Phospho-p53 (Ser46) Antibody.

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells treated with UV (100 J/m2 followed by a 3 hour recovery) and either Phospho-p53 (Ser15) Antibody or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CDKN1A Promoter Primers #6449, human MDM2 intron 2 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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货号：

37909T

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