Product Pathways - PathScan ELISA
PathScan® Acetylated p53 Sandwich ELISA Kit #7236
|7236S||1 Kit (96 assays)||---||In Stock||---|
|7236||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
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|Kit Includes||Volume||Solution Color|
|p53 Mouse mAb coated microwells||96 tests|
|Acetylated-Lysine Rabbit Detection Antibody||11 ml||Green|
|Anti-rabbit IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
|Human, Mouse, Monkey|
The PathScan® Acetylated p53 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of acetylated lysines on p53. A p53 Mouse monoclonal Antibody has been coated onto the microwells. After incubation with cell lysates, the p53 is captured by the coated antibody. Following extensive washing, Acetylated-Lysine Rabbit monoclonal Antibody is added to detect the acetylated lysines on the p53 protein. Anti-rabbit IgG, HRP linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of acetylated p53.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
CST's PathScan® Acetylated p53 Sandwich ELISA Kit detects endogenous levels of Acetylated p53. Using this Sandwich ELISA Kit #7236, acetylated lysines on p53 are detected when treated with TSA in COS cells. However, the levels of p53 remain unchanged, as shown by Western analysis (Figure 1). NIH/3T3 and 293 cells treated with TSA show similar results (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
ELISA - Western correlation
Figure 1: Treatment of COS cells with TSA causes accumulation of acetylation on p53, detected by Sandwich ELISA Kit #7236, but does not affect the level of p53 protein, detected by Western analysis. The absorbance at 450 nm is shown in the top figure, while the corresponding Western blot using the Acetylated Lysine mouse mAb (Ac-K-103) #9681 (left panel) or p53 Antibody #2524 (right panel), is shown in the bottom figure.
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).
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- Lohrum, M. and Scheidtmann, K.H. (1996) Oncogene 13, 2527-2539.
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- Solomon, J.M. et al. (2006) Mol. Cell. Biol. 26, 28-38.
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- 9441 Acetylated-Lysine Antibody
- 9681 Acetylated-Lysine Mouse mAb (Ac-K-103)
- 9814 Acetylated-Lysine (Ac-K2-100) Rabbit mAb
- 9282 p53 Antibody
- 2527 p53 (7F5) Rabbit mAb
- 2524 p53 (1C12) Mouse mAb
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 7004 TMB Substrate
- 7002 STOP Solution
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.