ab154470 is suitable to immunocapture p53 from whole cell lysates.
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- Cell reports 42:1120242023Protein adduction causes non-mutational inhibition of p53 tumor suppressor.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRavindran Caspa Gokulan,Kodisundaram Paulrasu,Jamal Azfar,Wael El-Rifai,Jianwen Que,Olivier G Boutaud,Yuguang Ban,Zhen Gao,Monica Garcia Buitrago,Sergey I Dikalov,Alexander I ZaikaPubMed 36848235
- The Journal of biological chemistry 288:36662-752013Resveratrol induces a mitochondrial complex I-dependent increase in NADH oxidation responsible for sirtuin activation in liver cells.
Key facts
- Sample type
- Tissue Extracts, Tissue Homogenate, Cell Lysate
- Reactive species
- Human
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application IP | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
Multifunctional transcription factor that induces cell cycle arrest, DNA repair or apoptosis upon binding to its target DNA sequence (PubMed:11025664, PubMed:12524540, PubMed:12810724, PubMed:15186775, PubMed:15340061, PubMed:17317671, PubMed:17349958, PubMed:19556538, PubMed:20673990, PubMed:20959462, PubMed:22726440, PubMed:24051492, PubMed:24652652, PubMed:35618207, PubMed:36634798, PubMed:38653238, PubMed:9840937). Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type (PubMed:11025664, PubMed:12524540, PubMed:12810724, PubMed:15186775, PubMed:15340061, PubMed:17189187, PubMed:17317671, PubMed:17349958, PubMed:19556538, PubMed:20673990, PubMed:20959462, PubMed:22726440, PubMed:24051492, PubMed:24652652, PubMed:38653238, PubMed:9840937). Negatively regulates cell division by controlling expression of a set of genes required for this process (PubMed:11025664, PubMed:12524540, PubMed:12810724, PubMed:15186775, PubMed:15340061, PubMed:17317671, PubMed:17349958, PubMed:19556538, PubMed:20673990, PubMed:20959462, PubMed:22726440, PubMed:24051492, PubMed:24652652, PubMed:9840937). 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 (PubMed:12524540, PubMed:17189187). Its pro-apoptotic activity is activated via its interaction with PPP1R13B/ASPP1 or TP53BP2/ASPP2 (PubMed:12524540). However, this activity is inhibited when the interaction with PPP1R13B/ASPP1 or TP53BP2/ASPP2 is displaced by PPP1R13L/iASPP (PubMed:12524540). In cooperation with mitochondrial PPIF is involved in activating oxidative stress-induced necrosis; the function is largely independent of transcription. Induces the transcription of long intergenic non-coding RNA p21 (lincRNA-p21) and lincRNA-Mkln1. LincRNA-p21 participates in TP53-dependent transcriptional repression leading to apoptosis and seems to have an effect on cell-cycle regulation. Implicated in Notch signaling cross-over. Prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression. 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. Regulates the circadian clock by repressing CLOCK-BMAL1-mediated transcriptional activation of PER2 (PubMed:24051492).
Alternative names
P53, TP53, Cellular tumor antigen p53, Antigen NY-CO-13, Phosphoprotein p53, Tumor suppressor p53
What's included?
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ab154470 is suitable to immunocapture p53 from whole cell lysates.
Key facts
- Sample type
- Tissue Extracts, Tissue Homogenate, Cell Lysate
- Reactive species
- Human
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
ab154470 is suitable to immunocapture p53 from whole cell lysates. Traditional immunoprecipitation methods usually result in co-elution of the antibody heavy and light chains that may co-migrate with relevant bands, masking important results. The abcam p53 immunocapture kit resolves this issue by immobilizing the p53 capture antibody onto protein G-agarose beads. The kit includes optimized buffers and reagents for sample preparation and p53 binding and recovery, which shorten the protocol and minimize handling and mixing.
p53 acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. It is 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 by p53 seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression. p53 is implicated in Notch signaling cross-over. p53 prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The protein p53 also known as TP53 or tumor protein p53 has a molecular weight of approximately 53 kDa. It acts as a transcription factor and plays a major role in cell cycle regulation apoptosis and maintaining genomic stability. This protein mainly expresses in the nucleus of cells and acts as a critical regulator of cellular responses to stress signals including DNA damage. Scientists commonly use p53 antibodies in various assays like western blot and p53 immunofluorescence to detect and study its expression and functional status in cells.
- Biological function summary
P53 functions to control cell division and apoptosis serving as a guardian of the genome by preventing mutation accumulation. It does not form part of a larger complex under normal conditions but interacts with various other molecules to execute its functions. p53 can activate or suppress the transcription of numerous genes involved in cell cycle arrest DNA repair and programmed cell death allowing it to halt the progression of damaged cells and trigger repair mechanisms or eliminate those that cannot be repaired.
- Pathways
P53 acts within several key biological pathways such as the p53 signaling pathway and the intrinsic apoptotic pathway. Its activity involves interaction with proteins like MDM2 which regulates p53 through ubiquitin-mediated degradation and ATM kinase which phosphorylates p53 in response to DNA damage. These interactions ensure appropriate cellular responses during stress and are vital for maintaining homeostasis.
- Associated diseases and disorders
P53 mutation or inactivation is often associated with the development of cancer given its role in controlling cell division and preventing tumor formation. Specifically its dysfunction has been linked to cancers such as breast cancer and lung cancer. Additionally p53 can interact with other mutant proteins like Ras compounding mutations that contribute to tumor progression and aggressive cancer phenotypes. Understanding these interactions and the status of p53 can be important in developing targeted cancer therapies.
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Immunoprecipitation - p53 Human Immunocapture Kit (ab154470)
Immunoprecipitation - p53 Human Immunocapture Kit (ab154470)
p53 immunocapture kit (ab154470) is specific to the p53 protein. Control proteins HSP60 and actin are present in the Hek293 and MCF7 lysate inputs, but are not found in p53 immunoprecipiations.
Hek293 cells were extracted with lauryl maltoside (lanes 2, 9), RIPA buffer (lanes 3, 10) or SDS (lanes 4, 11). MCF7 vehicle-treated cells were extracted with lauryl maltoside (lanes 5, 12) or RIPA buffer (lanes 7, 14). MCF7 cells treated with 1 μM camptothecin were extracted with lauryl maltoside (lanes 6, 13) or RIPA buffer (8, 15). Extracts of whole cells (20 μg, lanes 2-8), and one-fifth of immunoprecipitation samples using the p53 immunocapture beads (1 mg extract per 10 μL beads, lanes 9-15), were analyzed by Western blot using Anti-Hsp60 antibody - Mitochondrial Marker ab46798 anti-Hsp60 and Anti-ACTC1 antibody [EP184E] ab46805 anti-muscle actin.
Immunoprecipitation - p53 Human Immunocapture Kit (ab154470)
Western blot with ab154470 and MCF7 camptothecin-treated cells
All lanes: Anti-p53 antibody [E26] ab32389 anti-p53 1/1000
Lane 1: MW marker
Lane 2: MCF7 cells extracted with lauryl maltoside (LM).
Lane 3: MCF7 cells treated with 1 µM camptothecin for 6 hrs, extracted with LM.
Lane 4: MCF7 cells extracted with RIPA buffer.
Lane 5: MCF7 cells treated with 1 µM camptothecin for 6 hours, extracted with RIPA buffer.
Lane 6: IP with 10 µL p53 IP beads and 1 mg MCF7 cells, LM extract.
Lane 7: IP with 10 µL p53 IP beads and 1 mg MCF7 cells treated with 1 µM camptothecin for 6 hrs,
LM extract.
Lane 8: IP with 10 µL p53 IP beads and 1 mg MCF7 cells, RIPA extract.
Lane 9: IP with 10 µL p53 IP beads and 1 mg MCF7 cells treated with 1 µM camptothecin for 6 hrs,
RIPA extract.
Lane 10: IP with 10 µL Anti-GAL4 antibody [15-6E10A7] ab135397 beads and 1 mg MCF7 cells, RIPA extract.
Lane 11: IP with 10 µL Anti-GAL4 antibody [15-6E10A7] ab135397 beads and 1 mg MCF7 cells treated with 1 µM camptothecin for 6 hours, RIPA extract.
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