mouse anti human p53 antibody  (Proteintech)

Journal: PLOS One

Article Title: STAG2 mutations in the normal colon induce upregulation of oncogenic pathways in neighbouring wildtype cells

doi: 10.1371/journal.pone.0332499

Figure Lengend Snippet: Clusters 0 and 2 were comprised predominantly by wildtype cells. An upregulation of the “HALLMARK_TNFA_SIGNALING_VIA_NFKB” gene set in both Clusters 0 (a) and 2 (b) was observed. In Cluster 0, there was additionally an upregulation of “HALLMARK_ANGIOGENESIS” (c) and “HALLMARK_EPITHELIAL_MESENCHYMAL_TRANSITION” (d) . In Cluster 2, there was additionally an upregulation of “HALLMARK_EPITHELIAL_MESENCHYMAL_TRANSITION” (e) , “HALLMARK_KRAS_SIGNALING_UP” (f) , and “HALLMARK_P53_PATHWAY” (g) .

Article Snippet: Primary antibodies used included rabbit anti-human STAG2 antibody (1:100, 19837–1-AP, Proteintech, USA), mouse anti-human KI67 antibody (1:500, 66555–6-Ig, Proteintech, USA), mouse anti-human P53 antibody (1:400, 60283–2-Ig, Proteintech, USA), mouse anti-human CCND1 antibody (1:100, 60186–1-Ig, Proteintech, USA), mouse anti-human TERT antibody (1: 100, MA5−16033, Invitrogen, USA), mouse anti-human KRAS antibody (1:250, 415700, Invitrogen, USA), and mouse anti-human TNFα antibody (1:50, MA5−23720, Invitrogen, USA).

Techniques:

Journal: PLOS One

Article Title: STAG2 mutations in the normal colon induce upregulation of oncogenic pathways in neighbouring wildtype cells

doi: 10.1371/journal.pone.0332499

Figure Lengend Snippet: Fluorescent intensity of anti-TNFα antibody (a, b) , anti-KRAS antibody (c, d) and anti-p53 antibody (e, f) was statistically significantly upregulated in co-cultured wildtype organoids relative STAG2 mutant organoids. Scale bars are 50µm. All experiments were performed with N = 3 biological replicates.

Article Snippet: Primary antibodies used included rabbit anti-human STAG2 antibody (1:100, 19837–1-AP, Proteintech, USA), mouse anti-human KI67 antibody (1:500, 66555–6-Ig, Proteintech, USA), mouse anti-human P53 antibody (1:400, 60283–2-Ig, Proteintech, USA), mouse anti-human CCND1 antibody (1:100, 60186–1-Ig, Proteintech, USA), mouse anti-human TERT antibody (1: 100, MA5−16033, Invitrogen, USA), mouse anti-human KRAS antibody (1:250, 415700, Invitrogen, USA), and mouse anti-human TNFα antibody (1:50, MA5−23720, Invitrogen, USA).

Techniques: Cell Culture, Mutagenesis

Journal: PLOS One

Article Title: STAG2 mutations in the normal colon induce upregulation of oncogenic pathways in neighbouring wildtype cells

doi: 10.1371/journal.pone.0332499

Figure Lengend Snippet: In co-culture, we observed upregulation of TNFα, KRAS and p53 in wildtype organoids, proposing a cooperative mechanism of early oncogenesis.

Article Snippet: Primary antibodies used included rabbit anti-human STAG2 antibody (1:100, 19837–1-AP, Proteintech, USA), mouse anti-human KI67 antibody (1:500, 66555–6-Ig, Proteintech, USA), mouse anti-human P53 antibody (1:400, 60283–2-Ig, Proteintech, USA), mouse anti-human CCND1 antibody (1:100, 60186–1-Ig, Proteintech, USA), mouse anti-human TERT antibody (1: 100, MA5−16033, Invitrogen, USA), mouse anti-human KRAS antibody (1:250, 415700, Invitrogen, USA), and mouse anti-human TNFα antibody (1:50, MA5−23720, Invitrogen, USA).

Techniques: Co-Culture Assay

Journal: International Journal of Molecular Sciences

Article Title: Transcription Factor p73 Is a Predictor of Platinum Resistance and Promotes Aggressive Epithelial Ovarian Cancers

doi: 10.3390/ijms26073239

Figure Lengend Snippet: p73 and p53 protein expression in ovarian cancers. ( A1 ) Photomicrographs showing p73-negative ovarian cancers. ( A2 ) Photomicrographs showing cytoplasmic immunohistochemical staining of p73 in ovarian cancer tissue. ( A3 ) Photomicrographs showing nuclear immunohistochemical staining of p73 in ovarian cancer tissue. ( A4 ) Photomicrographs showing nuclear immunohistochemical staining of p53 in ovarian cancer tissue. ( B ) Kaplan–Meier curve for p73 nuclear/cytoplasmic co-expression and progression-free survival (PFS). ( C ) Kaplan–Meier curve for p53 nuclear protein expression and PFS. ( D ) Kaplan–Meier curve for p73/p53 co-expression and progression-free survival (PFS). Although 161 cases had expression data available for both p73 and p53 expression, clinical outcome data were available for only 147/161 patients and included in the survival analysis [light-blue line = p53−/p73−, aqua line = p53+/p73+, dark-red line = p53+/p73−, purple line = p53−/p73+]. SPSS software version 29 from IBM was used for Kaplan–Meier survival curves.

Article Snippet: Monoclonal mouse anti-human p53 [clone DO7, cell signalling] was used and was diluted at 1:100 in Leica antibody diluent (RE AR9352, Leica, Biosystems, Newcastle Upon Tyne, Tyne and Wear, UK) and incubated for 30 min at room temperature.

Techniques: Expressing, Immunohistochemical staining, Staining, Software

Journal: Scientific reports

Article Title: Extracorporeal shock waves effectively suppress colorectal cancer proliferation and growth.

doi: 10.1038/s41598-025-94386-3

Figure Lengend Snippet: Fig. 5. Effect of 60 mJ ESV on cell proliferation and death is related to p53 pathway. A Metascape analysis of upregulated genes in 60 mJ ESV group. B Gene set enrichment analysis plots for p53 pathway. C The relative mRNA level of p53 measured by real-time PCR. D Western blot analysis of P53. E P53 densitometry relative to β-actin. F P53 staining (Scale: 100 μm). Statistical significance was determined using one-way ANOVA with Tukey’s test. *p < 0.05.

Article Snippet: The membranes were probed overnight at 4 °C with primary antibody against human P53 (Cell Signaling Technology, 48818S, 1:1000), β-actin (Sigma, 1:10,000), cleaved-caspase3 (Cell Signaling Technology, 9664T, 1:1000), cleaved-PARP (Cell Signaling Technology, 9541T, 1:1000), AKR1C1 (Abcam, ab192785, 1:1000), COX2 (Abcam, ab179800, 1:1000), followed by incubation with peroxidase-conjugated secondary antibody (Cell Signaling Technology, 1:10,000) for 1.5 h. The signal was visualized with ECL (Millipore).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Staining

Journal: Scientific reports

Article Title: Extracorporeal shock waves effectively suppress colorectal cancer proliferation and growth.

doi: 10.1038/s41598-025-94386-3

Figure Lengend Snippet: Fig. 6.. 60 mJ ESV inhibits proliferation and promotes death of CRC cells in vitro. A HT29 cell viability at day 3 and day 6 after treatment with 60 mJ ESV, 2000 shots, 12 Hz. B Cells were analyzed by flow cytometry to determine the percentage of cells in the indicated cell cycle phases. C Cells were analyzed by flow cytometry to determine the percentage of death cells. D Western blot analysis of P53, cleaved-PARP, cleaved-caspase3, AKR1C1 and COX2. E P53, cleaved-PARP, cleaved-caspase3, AKR1C1 and COX2 densitometry relative to β-actin. Data are presented as means ± SD of three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s test. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The membranes were probed overnight at 4 °C with primary antibody against human P53 (Cell Signaling Technology, 48818S, 1:1000), β-actin (Sigma, 1:10,000), cleaved-caspase3 (Cell Signaling Technology, 9664T, 1:1000), cleaved-PARP (Cell Signaling Technology, 9541T, 1:1000), AKR1C1 (Abcam, ab192785, 1:1000), COX2 (Abcam, ab179800, 1:1000), followed by incubation with peroxidase-conjugated secondary antibody (Cell Signaling Technology, 1:10,000) for 1.5 h. The signal was visualized with ECL (Millipore).

Techniques: In Vitro, Flow Cytometry, Western Blot

Journal: Science Advances

Article Title: Nucleo-cytoplasmic environment modulates spatiotemporal p53 phase separation

doi: 10.1126/sciadv.ads0427

Figure Lengend Snippet: ( A ) Domain organization of p53 showing TAD (1 to 61), PRR (64 to 93), DBD (102 to 292), and TD (325 to 393) (top), low-complexity regions (blue) using SMART, disorder tendency (green) using IUPRED2A, residue-based droplet promoting probabilities (red), and droplet-promoting regions (DPR) (red box) using FuzDrop (bottom). ( B ) Structure of full-length p53 protein as predicted by Alphafold from 229 structures in PDB (UniProt ID P04637). ( C ) Confocal images of MCF10A cells showing p53 localization with (right) and without (left) cisplatin treatment. ( D ) The confocal microscopy showing the p53 (red) localization in MCF7 and MDA-MB-231. ( E ) STED microscopy of MDA-MB-231 and MCF7 cells showing p53 condensate state in the nucleus. Pseudocolor (LUT, mpl-plasma) has been used for representative purposes. ( F ) HeLa cells overexpressing WT GFP-p53 at 18 and 36 hours show cytoplasmic condensates. h, hours. ( G ) Time-lapse microscopic images showing fusion of WT GFP-p53 condensates in the cytoplasm of HeLa cells. ( H ) FRAP curves of cytoplasmic condensates in HeLa cells at 18 and 36 hours showing fluorescence recovery with (right) their corresponding t 1/2 . Note that the t 1/2 for condensates with low recovery was not estimated. ( I ) Time-dependent expression of WT GFP-p53 in SaOS2 cells showing nuclear (red arrowheads) and cytoplasmic condensates (white arrowheads). ( J ) Western blot images showing the expression of GFP-p53 with time in SaOS2 cells. ( K ) Time-dependent confocal images of cytoplasmic and nuclear condensates of WT GFP-p53 showing the fusion event of two condensates. ( L ) FRAP curves of nuclear (NC) and cytoplasmic condensate (CC) at 18 and 48 hours. All the experiments [(C) to (L)] were repeated two times with similar observations.

Article Snippet: Cells were then stained with mouse monoclonal anti-human p53 (DO-1) (catalog no. sc-126, Santa Cruz Biotechnology, USA) antibody (1:200) overnight at 4°C.

Techniques: Residue, Confocal Microscopy, Microscopy, Clinical Proteomics, Fluorescence, Expressing, Western Blot

Journal: Science Advances

Article Title: Nucleo-cytoplasmic environment modulates spatiotemporal p53 phase separation

doi: 10.1126/sciadv.ads0427

Figure Lengend Snippet: ( A ) Time-lapse confocal microscopy images showing spatiotemporal formation of p53 condensates. ( B ) Cytoplasmic/nuclear (C/N) ratio of WT GFP-p53 fluorescence signal in SaOS2 cells with time (mean ± SEM for n = 6 transfected cells). ( C ) Confocal microscopy images showing the effect of Leptomycin B (LMB) treatment (left) and corresponding C/N ratio calculation (right) of SaOS2 cells (mean ± SEM for n = 9 transfected cells). ( D ) The time-lapse confocal microscopy showing nuclear localization of the p53 with p53 NES− in SaOS2 cells. ( E ) STED microscopy of nuclear condensates of WT GFP-p53 at 18 and 48 hours. The line profile represents the intensity of GFP-p53 across the nucleus. ( F ) Lattice light-sheet imaging of nuclear condensates of SaOS2 transfected with WT GFP-p53 at 18 and 48 hours. Scale bars, 10 μm. ( G ) p53 condensates of R175H and R248Q in SaOS2 cells monitored over time using confocal microscopy. ( H ) FRAP recovery profiles of WT, R175H, and R248Q cytoplasmic condensates at 18 and 48 hours. ( I ) Size (area) and number distribution of nuclear condensates of WT, R175H, and R248Q at 18 and 48 hours, calculated from STED microscopy. Scale bars, 5 μm. ( J ) FRAP recovery profile of WT, R175H, and R248Q nuclear condensates at 18 and 48 hours. ( K ) Immunofluorescence study of WT GFP-p53 condensates with misfolded p53 specific antibody (Pab240). Scale bars, 10 μm. ( L ) Quantification of Pab240 colocalization with GFP-p53 nuclear and cytoplasmic condensates at 18 and 48 hours (mean with SEM, for N = 2). For (B), (C), and (L), the statistical significance was calculated using a two-tailed t test [*** P < 0.001, ** P < 0.002, * P < 0.033, and P > 0.012 (ns), with 95% confidence interval]. The experiments (A), (C) to (H), (J), and (K) were performed two independent times.

Article Snippet: Cells were then stained with mouse monoclonal anti-human p53 (DO-1) (catalog no. sc-126, Santa Cruz Biotechnology, USA) antibody (1:200) overnight at 4°C.

Techniques: Confocal Microscopy, Fluorescence, Transfection, Microscopy, Imaging, Immunofluorescence, Two Tailed Test

Journal: Science Advances

Article Title: Nucleo-cytoplasmic environment modulates spatiotemporal p53 phase separation

doi: 10.1126/sciadv.ads0427

Figure Lengend Snippet: ( A and B ) Western blot showing time-dependent expression of p21 (p53 target) and the corresponding fold change of p21. ( C ) Time-dependent apoptotic cell death of SaOS2 cells using FACS analysis. ( D ) p53 DNA binding capacity of p21 -specific DNA sequence by ELISA at 18 hours for WT and p53 mutants. ( E ) ChIP assay of p53 DNA binding to p21 -response element at 18 hours. For ELISA and ChIP assays, one-way analysis of variance (ANOVA) (*** P < 0.001, ** P < 0.01, * P < 0.05). ( F ) SaOS2 cells transfected with WT GFP-p53 after treatment with cisplatin showing p53 condensates at 18 hours (left). Scale bar, 10 μm. Super-resolution STED imaging showing the p53 clusters in the nucleus after treatment at 18 hours (right). ( G ) Size/number distribution of nuclear condensates for cisplatin-treated and untreated cells. ( H ) Condensates count in uniform area of (5 μm by 5 μm, N = 2) in the nucleus of cisplatin-treated SaOS2 cells. ( I and J ) Western blot of p53 expression with and without treatment of cisplatin (10 μM, 9 hours). ( K ) The p21 mRNA expression of SaOS2 cells at 18 hours in the presence and absence of cisplatin treatment {(H) and (K), two-tailed t test [*** P < 0.001, ** P < 0.002, * P < 0.033, and P > 0.012 (ns)]}, [(A) to (F) and (I) to (K)] N = 2. ( L ) Immunofluorescence of SaOS2 cells showing no colocalization with WT p53 and MDM2. In a few cells, cytoplasmic condensates of large size partially colocalizes with MDM2. ( M ) SaOS2 cells transfected with WT GFP-p53 after treatment with Nutlin showing p53 cytoplasmic condensates at 18 hours (left). Scale bar, 10 μm. STED imaging showing the presence of p53 clusters in the nucleus after treatment at 18 hours (right). ( N ) Estimation of condensate count in the nucleus of Nutlin-treated (Nut) and untreated (WT) cells. ( O ) FRAP recovery profiles of cytoplasmic condensates with Nutlin-treated cells.

Article Snippet: Cells were then stained with mouse monoclonal anti-human p53 (DO-1) (catalog no. sc-126, Santa Cruz Biotechnology, USA) antibody (1:200) overnight at 4°C.

Techniques: Western Blot, Expressing, Binding Assay, Sequencing, Enzyme-linked Immunosorbent Assay, Transfection, Imaging, Two Tailed Test, Immunofluorescence

Journal: Science Advances

Article Title: Nucleo-cytoplasmic environment modulates spatiotemporal p53 phase separation

doi: 10.1126/sciadv.ads0427

Figure Lengend Snippet: ( A ) The PDB structure of p53C (PDB ID: 2OCJ) shows random coil and β sheet conformation. ( B ) WT p53C shows disorder tendency (green), residue-based droplet-promoting probabilities (red), and droplet-promoting regions (DPR) (blue box). ( C ) Microscopy images showing the condensate formation of WT, R175H, and R248Q p53C ( N = 3). ( D ) Phase regimes (schematic) of WT, R175H, and R248Q. Opaque and open circles represent LLPS and no LLPS, respectively. ( E ) Static light scattering (LS at 350 nm) showing condensate formation by WT p53C, R175H, and R248Q in the presence (filled circles) and absence (open circles) of 10% PEG-8000. ( F ) Time-lapse DIC images showing the fusion event of two WT p53 condensates with time. Scale bars, 2 μm. ( G ) FRAP experiment at different time intervals of WT p53C, R175H, and R248Q variants with t 1/2 calculation (right). ( H ) FTIR spectroscopy of dense and dilute phase of WT p53 after centrifugation. ( I ) Confocal microscopy images WT p53C condensates at different time intervals. ( J ) The size distribution of WT, R175H, and R248Q condensates with time. ( K ) TEM micrographs WT p53C, R175H, and R248Q condensates at 0 and 12 hours. ( L ) The ThT binding p53C condensates at 12 hours. α-Synuclein fibrils were used as a positive control. N = 3 for (C), (D), and (I), and N = 2 for (E) to (H), (K), and (L). ( M ) ThioS staining of WT p53C condensates formed in the presence and absence of CSA at indicated time points ( n = 3 independent experiments). ( N ) The FRAP of WT p53C condensates in the presence of CSA. ( O ) The ThT binding for p53C condensates in the presence of CSA. α-Synuclein fibrils were used as a positive control ( N = 3). ( P ) TEM micrographs of condensates in the presence of CSA.

Article Snippet: Cells were then stained with mouse monoclonal anti-human p53 (DO-1) (catalog no. sc-126, Santa Cruz Biotechnology, USA) antibody (1:200) overnight at 4°C.

Techniques: Residue, Microscopy, Spectroscopy, Centrifugation, Confocal Microscopy, Binding Assay, Positive Control, Staining

Journal: Science Advances

Article Title: Nucleo-cytoplasmic environment modulates spatiotemporal p53 phase separation

doi: 10.1126/sciadv.ads0427

Figure Lengend Snippet: ( A ) Confocal images of WT p53C condensates formed in the presence and absence of target DNA (TR-DNA), nontarget DNA (NTR-DNA), and RNA. The inset for NTR-DNA and TR-DNA represents multicomponent condensate formation. ( B ) Size (Feret) distributions of p53C condensates formed in the presence and absence of TR-DNA, NTR-DNA, and RNA. ( C and D ) Phase regime of WT p53C with varying concentrations of NTR-DNA and RNA. Opaque and open circles represent LLPS and no LLPS, respectively. Pink shaded region showing p53 condensates in the absence of any nucleic acid. ( E ) Static light scattering (LS at 350 nm) showing the effect of RNA, NTR-DNA, and TR-DNA for p53C condensate formation. ( F ) FRAP profile of p53C condensates in the presence of NTR-DNA and RNA showing a complete recovery at the early (0 hours) and late (12 hours) time points, suggesting maintenance of the liquid-like state. ( G ) The confocal image of NHS-Rhodamine–labeled p53C condensates formed in the cellular fraction of the nuclear extract (NE) and cytoplasmic extract (CE). Scale bar, 2 μm. ( H ) The size and circularity distribution of p53C condensates formed in the presence of CE and NE showing larger size in CE with lesser circularity in comparison to condensate formed in the NE. ( I ) FRAP of p53C condensates formed in NE and CE. The experiments (A), (C), (D), and (G) were repeated three independent times and (E), (F), and (I) were repeated two independent times. ( J ) The confocal image of p53C mutant (R175H and R248Q) condensates formed in NE (left) . FRAP profile of R175H and R248Q condensates formed in NE (right). ( K ) Representative confocal images of preformed R248Q condensates after adding TR-DNA and NTR-DNA. ( L ) FRAP of preformed R248Q condensates after adding TR-DNA and NTR-DNA, showing negligible fluorescence recovery.

Article Snippet: Cells were then stained with mouse monoclonal anti-human p53 (DO-1) (catalog no. sc-126, Santa Cruz Biotechnology, USA) antibody (1:200) overnight at 4°C.

Techniques: Labeling, Comparison, Mutagenesis, Fluorescence

Journal: Science Advances

Article Title: Nucleo-cytoplasmic environment modulates spatiotemporal p53 phase separation

doi: 10.1126/sciadv.ads0427

Figure Lengend Snippet: ( A ) Schematic showing the possible mode of operation of p53 in the nuclear microenvironment. ( B ) Left: Representative images of preformed p53 condensates after the addition onto the uncoated and TR- and NTR-DNA-coated coverslips. Right: The calculated number of condensates with various nucleic acid–coated surfaces and uncoated coverslips were taken as control (C). The statistical significance was determined by one-way ANOVA using Newman-Keuls (SNK) post hoc analysis (*** P < 0.001, ** P < 0.01). ( C ) Split-violin plot represents the change in the partitioning of p53C after adding TR-DNA and NTR-DNA. ( D ) Static light scattering (LS at 350 nm) showing the kinetics of p53C in the presence of 10% (w/v) PEG-8000 and the effect after the addition of TR-DNA, NTR-DNA, and RNA. The data showing a decrease in light scattering value in the presence of TR-DNA, suggesting the dissolution of preformed p53C condensates. ( E ) The confocal microscopy images showing the effect of TR-DNA on preformed p53C condensates formed in the presence of NTR-DNA (p53C-NTR DNA), RNA (P53C-RNA), and RNA + NTR-DNA (p53-RNA-NTR-DNA) condensates. ( F to H ) Size distribution plot showing the reduction in the size/numbers of condensates after the addition of TR-DNA to (F) p53C-RNA condensates, (G) p53C-NTR-DNA condensates, and (H) p53C-RNA-NTR-DNA condensates. ( I to K ) The confocal microscopy images show few residual condensates observed after condensate dissolution due to the addition of TR-DNA. The left panel represents Rhod-p53 condensates formed in the presence of (I) Atto 488–labeled NTR DNA, (J) RNA, and (K) RNA and Atto 488–labeled NTR-DNA. The middle panel represents residual p53C condensates that remained after the addition of Atto 647N–labeled TR-DNA. The right panel represents a line profile across the condensates to show the spatial distribution of protein and DNA. The experiments (B), (D), (E), and (I) were repeated two independent times.

Article Snippet: Cells were then stained with mouse monoclonal anti-human p53 (DO-1) (catalog no. sc-126, Santa Cruz Biotechnology, USA) antibody (1:200) overnight at 4°C.

Techniques: Control, Dissolution, Confocal Microscopy, Labeling

Journal: Science Advances

Article Title: Nucleo-cytoplasmic environment modulates spatiotemporal p53 phase separation

doi: 10.1126/sciadv.ads0427

Figure Lengend Snippet: ( A ) The proposed model of p53 regulation through p53 phase separation. Crystal structure showing the core domain bound with DNA consensus element of p53 in tetrameric form. ( B ) Dynamic light scattering profile of p53C monomer, p53C condensates alone, and p53C condensates dissolved in the presence of the specific DNA. ( C ) SEC measurement of p53C monomer and p53C condensate in the presence of target DNA showing a peak approximately at 100 kDa, confirming tetrameric state. γ-Globulin (150 kDa), lactoferrin (78 kDa), and chymotrypsin (25 kDa) are used as markers in SEC. ( D ) The SEC eluted the oligomeric fraction of p53C condensates in the presence of TR-DNA, showing the presence of DNA bound with protein using UV spectroscopy (left) and agarose gel electrophoresis (right). ( E ) Spectral shift measurement of p53C in the presence of TR-DNA showing binding affinity of p53C with nucleic acids. ( F ) Bar graph representing the binding affinity ( K d ) of p53C with TR-DNA and NTR-DNA showing a significantly higher binding affinity for TR-DNA. Error bars represent mean ± SEM for N = 3 independent experiments. The statistical significance was estimated with an unpaired t test with 95% confidence interval (** P < 0.002). ( G ) The change in molar ellipticity at θ 222 with an increase in temperature shows no major changes in temperature denaturing profiles between the LLPS and non-LLPS state of p53C. Representative microscopic images of p53C in LLPS and non-LLPS states for the temperature-dependent CD experiment are shown. ( H ) Proteinase K (PK) digestion was followed by SDS-PAGE and showed the higher PK resistivity of the p53 LLPS state. ( I ) SDS-PAGE quantification of PK digestion assay of LLPS and non-LLPS p53C showing faster PK digestion with time of p53C in the non-LLPS state compared to the LLPS state. The experiments (B) to (H) were repeated two independent times.

Article Snippet: Cells were then stained with mouse monoclonal anti-human p53 (DO-1) (catalog no. sc-126, Santa Cruz Biotechnology, USA) antibody (1:200) overnight at 4°C.

Techniques: Spectroscopy, Agarose Gel Electrophoresis, Binding Assay, SDS Page