P53 do 7

Manufactured by Agilent Technologies

Sourced in United States

The P53 (DO-7) is a laboratory equipment product offered by Agilent Technologies. It is used for the detection and quantification of the p53 protein, a key tumor suppressor. The device functions to measure p53 levels in biological samples, providing researchers with important data for understanding cellular processes and disease states.

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Lab products found in correlation

Progesterone receptor 1e2, by Roche (1 mentions) Ki 67 mib 1, by Roche (1 mentions) Ventana autostainer, by Roche (1 mentions) Kit cd117, by Agilent Technologies (1 mentions) Desmin d33, by Agilent Technologies (1 mentions) Chromogranin a, by Nichirei Biosciences (1 mentions) Odyssey software, by LI COR (1 mentions) Irdye 800cw donkey anti rabbit igg, by LI COR (1 mentions) Phospho h2ax ser139, by Merck Group (1 mentions) Gapdh, by Novus Biologicals (1 mentions) Irdye 800cw donkey anti mouse igg, by LI COR (1 mentions) Alexa fluor 680 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 680 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Odyssey imager, by LI COR (1 mentions) Phospho p53 ser15, by Cell Signaling Technology (1 mentions) Cxcr4, by Abcam (1 mentions) Bcl2 clone 124, by Agilent Technologies (1 mentions) Cd30 clone berh2, by Agilent Technologies (1 mentions) Irf4 mum1, by Agilent Technologies (1 mentions) Foxp1, by Abcam (1 mentions)

11 protocols using p53 do 7

Immunohistochemical Profiling of Tissue Microarray

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Serial sections of the TMA were stained with protein marker in immunohistochemistry either using the Ventana autostainer (Roche), or manually, following manufacturers’ protocols. The following antibodies were used: Estrogen Receptor (SP1, Ventana Medical Systems), Progesterone Receptor (1E2, Ventana Medical Systems), HER2 (4B5, Ventana Medical Systems), Ki67 (MIB-1, Ventana Medical Systems), P53 (DO-7, Dako), P16 (CINtec, Ventana Medical Systems), P21 (1:200, Cell Signaling Cat#2947). ER, PR, HER2 and P21 IHC were scored by study pathologist (K.L.) with an evaluation of staining intensity and percentage of positive cells [28 (link)]. HER2 scoring was conducted according to the recommendations from 2013 ASCO/CAP [29 (link)]. HER2 2+ (equivocal) cases by IHC were further evaluated with fluorescent in situ hybridization (FISH) (S. N-M). Ki67 IHC was presented as percentages of positive cells per core (scored by K.L.). P53 and P16 IHC were reviewed by study pathologist (S. N-M.) and categorized as normal, null or over-expressed for P53 and normal or over-expressed for P16.

Cheung A.M., Wang D., Liu K., Hope T., Murray M., Ginty F., Nofech-Mozes S., Martel A.L, & Yaffe M.J. (2021). Quantitative single-cell analysis of immunofluorescence protein multiplex images illustrates biomarker spatial heterogeneity within breast cancer subtypes. Breast Cancer Research : BCR, 23, 114.

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Histological Diagnosis of GI-SELs via FNA-CB and Resected Specimens

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All histocytological diagnoses for FNA-CB specimens and resected specimens were prospectively made via the consensus of two or more pathologists (YN, TS, MU, and FF). To assess the histology of the FNA-CB specimens, cell-block sections were prepared using a sodium alginate method and subjected to hematoxylin and eosin (HE) stain. Immunostaining was performed when the following situations occurred: 1) GI-SELs for which immunostaining would be diagnostic (e.g., GISTs, leiomyoma, schwannoma, etc.) were suspected based on HE staining, and 2) HE staining afforded an indeterminate histological diagnosis. The antibodies used for immunostaining were as follows: KIT (CD117; DAKO, Glostrup, Denmark), CD34 (NU-4A1; Nichirei, Tokyo, Japan), DOG1 (K9; Novocastra, Newcastle, UK), Desmin (D33; DAKO), αSMA (1A4; Enzo Life Sciences, Farmingdale, USA), Ki67 (MIB-1; Immunotech, Marseilles, France), P53 (DO-7; DAKO), MUC1 (Ma695; Novocastra), chromogranin A (chromogranin; Nichirei), and synaptophysin (27G12; Novocastra).
For resected specimens, HE staining and several immunostainings were performed using the method for FNA-CB specimens. When histological diagnoses for resected specimens could be made using HE staining, immunostaining was not performed.

Shimizu T., Koshita S., Ohira T., Harada Y., Kanno Y., Ogawa T., Yamagata T., Kusunose H., Sakai T., Tsuchiya T., Oikawa M., Noda Y., Sawai T, & Ito K. (2021). Endoscopic Ultrasonography-guided Fine-needle Aspiration Cytology Combined with a Cell-block Method for Gastrointestinal Subepithelial Lesions. Internal Medicine, 61(7), 935-942.

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Western Blot Analysis of DNA Damage Markers

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Protein lysates were extracted using UTB (9 M urea, 0.75 M Tris–HCL [pH 7.5] and 0.15 M β-mercaptoethanol) and WB was performed as previously described^{24 (link)}. Primary antibodies used were p53 DO-7 (DAKO, #M7001), phospho-p53 (ser15) (Cell signalling, #9284), phospho-H2AX (ser139) (Millipore, #05-636, clone JBW301), H2AX (Millipore, AB10022), p21 (Cell signalling, #2947), GAPDH (Novus Bio, #NB600-502). Secondary antibodies used were Alexa-Fluor 680 goat anti-mouse IgG (Invitrogen), Alexa-Fluor 680 goat anti-rabbit IgG (Invitrogen), IRDye 800CW donkey anti-mouse IgG (LI-COR) and IRDye 800CW donkey anti-rabbit IgG (LI-COR). All membranes were scanned using the Odyssey Imager and images acquired using Odyssey software (LI-COR Biosciences, Lincoln NE).

Anbalagan S., Ström C., Downs J.A., Jeggo P.A., McBay D., Wilkins A., Rothkamm K., Harrington K.J., Yarnold J.R, & Somaiah N. (2021). TP53 modulates radiotherapy fraction size sensitivity in normal and malignant cells. Scientific Reports, 11, 7119.

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Prognostic Biomarkers in Diffuse Large B-Cell Lymphoma

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Tissue microarrays prepared from the diagnostic formalin-fixed, paraffin-embedded (FFPE) tissue blocks of all patients studied were stained with an anti-p63 antibody (4A4, Santa Cruz Biotechnology, Santa Cruz, CA) which can detect all p63 isoforms. Expression levels of p63 were determined by estimating the percentage of p63-positive tumor cells in the tissue array cores. X-tile software and receiver operating characteristic curve analysis by GraphPad Prism 6 Software were used to determine the percentage of p63-positive cells with maximal discriminatory power for the separation of DLBCL patients into 2 different prognostic groups. Evaluation of other biomarkers by immunohistochemistry was also performed on tissue microarrays using corresponding antibodies: p53 (DO-7, Dako, Carpinteria, CA), MDM2 (IF2, Calbiochem, Billerica, MA), p21 (Dako), Bcl-2 (Clone-124, Dako, Carpinteria, CA), Ki-67 (Dako), CD30 (clone BerH2, Dako), Bcl-6 (Dako), FOXP1 (Abcam), IRF4/MUM1 (Dako), CD10 (56C6, Vantana), c-Rel (Dako), and CXCR4 (Abcam, San Francisco, CA). Details of immunohistochemistry procedures and scoring processes have been described previously [38 (link), 44 (link), 55 (link)–58 (link)].

Xu-Monette Z.Y., Zhang S., Li X., Manyam G.C., Wang X.X., Xia Y., Visco C., Tzankov A., Zhang L., Montes-Moreno S., Dybkaer K., Chiu A., Orazi A., Zu Y., Bhagat G., Richards K.L., Hsi E.D., Choi W.W., van Krieken J.H., Huh J., Ponzoni M., Ferreri A.J., Zhao X., Møller M.B., Parsons B.M., Winter J.N., Piris M.A., Medeiros L.J, & Young K.H. (2016). p63 expression confers significantly better survival outcomes in high-risk diffuse large B-cell lymphoma and demonstrates p53-like and p53-independent tumor suppressor function. Aging (Albany NY), 8(2), 345-365.

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Immunohistochemical Analysis of Signaling Proteins

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IHC procedures were described previously [17 (link)]. Antibodies used were Wip1 (F-10) (mouse monoclonal, 1:100, Santa Cruz, Dallas, TX, USA), p53 (DO-7) (mouse monoclonal, 1:200, Dako, Tokyo, Japan), phospho-p53 (S15) (rabbit polyclonal, 1:1000, Abcam, Cambridge, UK), and phospho-p38 MAPK (Thr180/Tyr182) (D3F9) (rabbit monoclonal, 1:1000, Cell Signaling, Danvers, MA, USA). The representative images of staining are displayed in Fig. 1 [17 (link)].Fig. 1

Representative images for immunohistochemical evaluations. The 0 to 3 staining intensity of Wip1, nuclear phospho-p38, cytoplasmic phospho-p38, and phospho-p53 as well as the percentages of nuclear p53 staining. ×200

Xu C., Minaguchi T., Qi N., Fujieda K., Suto A., Itagaki H., Shikama A., Tasaka N., Akiyama A., Nakao S., Ochi H, & Satoh T. (2022). Differential roles of the Wip1–p38–p53 DNA damage response pathway in early/advanced-stage ovarian clear cell carcinomas. World Journal of Surgical Oncology, 20, 139.

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Protein Expression Analysis by Western Blot

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Protein isolation and western blot analysis were performed as described previously [17 (link)]. Primary antibodies used were p53 (DO-7, Dako, CA, USA), p21 (c-19, Santa Cruz), ANXA A1, ANXA A2 (Becton Dickinson) and β-actin (Abcam), followed by mouse secondary conjugated antibody (Abcam). Protein abundance was quantified by image analysis using the Kodak image station 4000MM.

Hatoum D., Yagoub D., Ahadi A., Nassif N.T, & McGowan E.M. (2017). Annexin/S100A Protein Family Regulation through p14ARF-p53 Activation: A Role in Cell Survival and Predicting Treatment Outcomes in Breast Cancer. PLoS ONE, 12(1), e0169925.

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Immunohistochemical Analysis of Cell Proliferation

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Raft sections were deparaffinized and dehydrated in xylene and alcohol sequential baths. The endogenous peroxidase activity was quenched with 3% H₂O₂ after antigen retrieval in boiling citrate buffer. Primary antibodies against BrdU (1:400; Zymed, San Francisco, CA, USA), p53 DO-7 (1:400; DakoCytomation, Glostrup, Denmark) and pRb (1:500; Novocastra, Newcastle-upon-Tyne, UK) were incubated for 18 h in 1% bovine serum albumin-phosphate buffered solution. After incubation with secondary antibody, antigens were detected with streptavidin-biotin-peroxidase complex (StreptABComplex/HRP Duet Mouse/Rabbit DakoCytomation, Dako). Chromogenic detection of peroxidase was performed with diaminobenzidine (DAB) substrate (Sigma-Aldrich, St Louis, MO, USA). The percentage of BrdU-positive/total nuclei was determined by direct counting cell nuclei. At least 3000 nuclei were counted per experiment.

Rabachini T., Boccardo E., Andrade R., Perez K.R., Nonogaki S., Cuccovia I.M, & Villa L.L. (2018). HPV-16 E7 expression up-regulates phospholipase D activity and promotes rapamycin resistance in a pRB-dependent manner. BMC Cancer, 18, 485.

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Immunoblotting Antibodies and Targets

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Immunoblotting was performed as described previously [18 (link)]. The following antibodies were used: α-tubulin and β-actin (Sigma-Aldrich), p21^Cip1/WAF1 and p27^KIP1 (BD-Pharmingen, San Diego, CA); p53 (DO-7; Dako, Carpinteria, CA); BRCA1(Santa Cruz Biotechnology, Dallas, TX); Cdc2 (MBL, Nagoya, Japan); PIM-1 and XPO1 (marketed as anti-CRM1) and p-HSF1^Ser326 (Abcam, Cambridge, MA); CDC25C, c-Myc, cyclin D1, Hsp70, 4E-BP1, phosphorylated- (p-) 4E-BP1^Thr37/Thr46, p-Rb^Ser780, S6 (S6K), p-S6 ribosomal protein (S6K)^{Ser235/Ser236}, PUMA, HSF1, Hsp70 and horseradish peroxidase–linked anti-mouse and anti-rabbit IgG (all from Cell Signaling Technology).

Tabe Y., Kojima K., Yamamoto S., Sekihara K., Matsushita H., Davis R.E., Wang Z., Ma W., Ishizawa J., Kazuno S., Kauffman M., Shacham S., Fujimura T., Ueno T., Miida T, & Andreeff M. (2015). Ribosomal Biogenesis and Translational Flux Inhibition by the Selective Inhibitor of Nuclear Export (SINE) XPO1 Antagonist KPT-185. PLoS ONE, 10(9), e0137210.

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Immunohistochemical Profiling of FFPE Tumor Tissue

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Tumour tissue and the agarose-embedded close-to-patient cancer cells were formalin fixed and paraffin embedded (FFPE) before 4 μm sections were cut for both H&E and immunohistochemistry (IHC) analysis. IHC was performed using standard techniques and in line with the manufacturer's instructions for the following primary antibodies: Cytokeratin (MNF116, DAKO), EpCam (Ber-EP4, DAKO), CD44 (DF1485, DAKO), p53 (DO-7, DAKO), Vimentin (V9, DAKO), TFF3 (Abcam), and ALDH1A1 (EP1933Y, Abcam) (see online supplementary method S6). Sections were viewed with a Leica DMLB Bright-field Microscope (Leica-microsystems, Milton Keynes, UK) and images acquired with Leica QWin Standard v3 software. The presence of any characteristically stained cells was considered positive with respect to negative controls, and confirmed by a second blinded individual.

Saunders J.H., Onion D., Collier P., Dorrington M.S., Argent R.H., Clarke P.A., Reece-Smith A.M., Parsons S.L, & Grabowska A.M. (2016). Individual patient oesophageal cancer 3D models for tailored treatment. Oncotarget, 8(15), 24224-24236.

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Immunohistochemical Profiling of Soft Tissue

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Immunohistochemistry was performed on 4 μm thick formalin-fixed paraffin-embedded (FFPE) tissue for pan-TRK (clone EPR17341, 1:300, Abcam, Cambridge, MA), S100 (polyclonal, 1:3000, Dako, Carpinteria, CA), SOX10 (clone EP268, 1:2000, Cell Marque, Rocklin, CA), CD34 (clone M7165, 1:150, Dako), desmin (clone DE-U-10, 1:5000, Sigma-Aldrich, Saint Louis, MO), smooth muscle actin (SMA, clone 1A4, 1:20 000, Sigma), h-caldesmon (clone h-CD, 1:300, Dako), estrogen receptor (ER, clone SP1, 1:40, Fisher Scientific, Hampton, NH), progesterone receptor (PR, PgR636, 1:200, Dako), and p53 (DO-7, 1:500, Dako).

Costigan D.C., Nucci M.R., Dickson B.C., Chang M.C., Song S., Sholl L.M., Hornick J.L., Fletcher C.D, & Kolin D.L. (2022). NTRK-Rearranged Uterine Sarcomas: Clinicopathologic Features of 15 Cases, Literature Review, and Risk Stratification. The American journal of surgical pathology.

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