Western blot assays were performed using whole liver lysates 41 (link) or nuclear proteins of the liver samples and images were collected by ImageQuant LAS 4000 (GE Healthcare, Pittsburgh, PA) 42 (link). MTP antibody (catalog sc-135994), HNF4α antibody (catalog sc-6556) and p53 antibody (catalog sc-6243) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). ApoB antibody was purchased from Meridian Life Science (K45253G, TN). β-actin antibody was from Novus Biologicals (catalog NB600-501, CO). Histone antibody was from Cell Signaling (Beverly, MA). The antibodies were used at a concentration of 1 μg per ml.
P53 antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The P53 antibody is a laboratory reagent used in various applications, including Western blotting, immunohistochemistry, and immunocytochemistry. It is designed to specifically detect the p53 protein, a tumor suppressor that plays a crucial role in cell cycle regulation and apoptosis.
Automatically generated - may contain errors
Lab products found in correlation
Mtp antibody, by Santa Cruz Biotechnology (2 mentions)
Hnf4α antibody, by Santa Cruz Biotechnology (2 mentions)
Apob antibody, by Meridian Bioscience (2 mentions)
β actin antibody, by Novus Biologicals (2 mentions)
Histone antibody, by Cell Signaling Technology (2 mentions)
Imagequant las 4000, by GE Healthcare (2 mentions)
Anti hsp90 antibody, by Cell Signaling Technology (2 mentions)
Universal magnetic co ip kit, by Active Motif (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Normal rabbit igg, by Merck Group (1 mentions)
Chip kit, by Merck Group (1 mentions)
Phospho p53 antibodies, by Cell Signaling Technology (1 mentions)
Cxcr4 antibody, by Boster Bio (1 mentions)
Keratinocyte serum free medium, by Thermo Fisher Scientific (1 mentions)
N n hexamethylenebisacrylamide hmba, by Polysciences (1 mentions)
Cyclam, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Hdac9 antibody, by Abcam (1 mentions)
Trps1 antibody, by R&D Systems (1 mentions)
Anti mouse secondary antibody, by Proteintech (1 mentions)
16 protocols using p53 antibody
1
Quantitative Protein Analysis in Liver
2
ChIP Assay for p53 Binding in Drp1 Promoter
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Chromatin immunoprecipitation (ChIP) was carried out with a ChIP kit (Millipore, Billerica, MA) according to manufacturer's instructions. Briefly, chromatin was cross-linked by formaldehyde for 10 min at room temperature. Crosslinking reaction was quenched by glycinesolution for 5 min. Cross-linked chromatin was sonicated into fragments at 4°C. The purified chromatin was immunoprecipitated with p53 antibody (Santa Cruz Biotechnology) or normal rabbit IgG (Millipore). DNA/Protein complexes were collected by the protein G-agarose beads, which were washed and bound DNA was eluted. The eluted DNA was used in PCR amplification with the primers which encompass p53 binding site (BS)1 or BS2 of the rat Drp1 promoter. The primers were as follows: BS1 (corresponding to a 225 bp fragment), Forward: 5′-GGC TGT ATG TGT TCC ATT AC-3′; Reverse: 5′-AGA CAG AAG AGA GTA GGC TC-3′. BS2 (corresponding to a 220 bp fragment), Forward: 5′-AGT AAA GCC TGT CTT GTG TG-39; Reverse: 5′-AAA TAA TCA CAA TAT ACT G-3′.
3
Immunoblot Analysis of p53 Signaling
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Immunoblot was performed as described in detail [74 (link)]. The p53 antibody was from Santa Cruz (Dallas, TX, USA). Phospho-p53 antibodies were ordered from Cell Signaling (Frankfurt am Main, Germany). Quantification of immunoblots was done as described [75 (link)].
4
Optimizing HMBA-Mediated Gene Delivery
5
ChIP Assay for DNA-Protein Interactions
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ChIP assays were performed as previously described [54 (link)]. Briefly, etoposide- or H2O2-treated U2OS, HeLa, and SH-SY5Y cells were harvested and cross-linked with 1% formaldehyde. The DNA-protein complex was precipitated by the p53 antibody (Santa Cruz, USA). The primers used to detect immunoprecipitated DNA are shown in Supplementary Table 1 .
6
Antibody-based Detection of Histone Modifications
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Antibodies for HDAC1–6 and 53BP1 were obtained from Cell Signaling Technology (Beverly, MA). TRPS1 antibody was purchased from R&D Systems (Minneapolis, MN). HDAC9 antibody was purchased from Abcam (Cambridge, MA). H3 and H4 antibodies were obtained from EMD Millipore (Billerica, MA). Ac-H3K9, ac-H3K18 and ac-H4K16 antibodies were purchased from Active Motif (Carlsbad, CA). P53 antibody and anti-rabbit and anti-goat secondary antibodies were purchased from Santa Cruz Biotechnology. β-actin antibody and anti-mouse secondary antibody were purchased from Proteintech (Chicago, IL). For western blotting, protein lysates were separated by SDS-PAGE, transferred to PVDF membranes, and immunoblotted with the respective antibodies as indicated above and in the figures. Blots were developed with SuperSignal West Femto Maximum Sensitivity Substrate (Pierce/Thermo Scientific, Rockford, IL).
7
ChIP Assay Protocol with Antibodies
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ChIP assays were performed using EZ-CHIP KIT according to the manufacturer's instruction (Millipore). p53 antibody was from Santa Cruz Biotechnology. NF-YA antibody was obtained from Abcam. The ChIP primer sequences were listed in Supplementary Table S1 . Quantification of immunoprecipitated DNA was performed using qPCR with SYBR Green Mix (Takara). ChIP data were calculated as a percentage relative to the input DNA by the equation 2[Input Ct- Target Ct] × 0.1 × 100.
8
Western Blot Analysis of Protein Expression
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Cells were harvested and resuspended in cell lysis buffer (150 mM NaCl, 50 mM HEPES (pH 7.5), 1% NP40) containing a protease inhibitor cocktail (Roche, Basel, Switzerland). Whole-cell lysates were resolved by SDS-PAGE and transferred to PVDF membranes (GE Healthcare, Uppsala, Sweden) via Western blotting. Proteins were detected with a 1:1000 or 1:5000 dilution of the primary antibody using a chemiluminescence system (Dogen, Seoul, Korea). Images were acquired using the LAS4000 system (GE Healthcare, Uppsala, Sweden). The CTRP1 antibody was purchased from Invitrogen (Carlsbad, CA, USA), p21 antibody from Cell Signaling Technology (Danvers, MA, USA), and p53 antibody from Santa Cruz Biotechnology (Dallas, TX, USA). FR180204, ERK inhibitor, was purchased from Calbiochem (San Diego, CA, USA).
9
Quantitative Protein Analysis in Liver
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Western blot assays were performed using whole liver lysates 41 (link) or nuclear proteins of the liver samples and images were collected by ImageQuant LAS 4000 (GE Healthcare, Pittsburgh, PA) 42 (link). MTP antibody (catalog sc-135994), HNF4α antibody (catalog sc-6556) and p53 antibody (catalog sc-6243) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). ApoB antibody was purchased from Meridian Life Science (K45253G, TN). β-actin antibody was from Novus Biologicals (catalog NB600-501, CO). Histone antibody was from Cell Signaling (Beverly, MA). The antibodies were used at a concentration of 1 μg per ml.
10
Co-immunoprecipitation of p53 and HSP90
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Co-immunoprecipitation was carried out using a Universal Magnetic Co-IP kit (Active Motif, Cat. # 54002). Small EVs from GOF TP53 HT29 and wt TP53 RKO cells were isolated as described above. Small EVs were lysed using kit lysis buffer, and 5 μg of p53 antibody (Santa Cruz, Cat. # sc-126) per sample was used for the pull-down of p53 and HSP90. The resulting proteins immunoprecipitated with p53 were then run on a denaturing gel, as described above. The presence of immunoprecipitated HSP90 in the same samples was determined by probing the blot with an anti-HSP90 antibody (Cell Signaling Technology, Cat. # 4877, 1:1000).
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