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P53 polyclonal antibody fl393

Manufactured by Santa Cruz Biotechnology

The P53 polyclonal antibody (FL393) is a laboratory reagent produced by Santa Cruz Biotechnology. It is a purified rabbit polyclonal antibody that recognizes the full-length p53 protein. The antibody can be used for the detection of p53 in various applications, such as western blotting, immunoprecipitation, and immunohistochemistry.

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3 protocols using p53 polyclonal antibody fl393

1

Plasmid Constructs for p53 Regulation

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Plasmid of pGL4-p53 was purchased from Promega. The Plasmids of pCDNA3.1-CDR1as was a gift of Dr. Nikolaus Rajewsky. Plasmids of Myc-p53, Myc-MDM2, PCDH-p53 and HA-Ub were a gift of Dr. Zhang Lingqiang. The pCDNA3.1(+) circRNA mini vector was purchased from Addgene (60648). The plasmid of HA-MDM2 was purchased from Sino Biological (HG11206-CY). All expression constructs were verified by DNA Sequencing.
Antibodies against p53 (DO-1) (ab1101), MDM2 (ab3110), or PUMA (ab33906) were purchased from Abcam Inc. The p53 polyclonal antibody (FL393) and the Myc antibody (9E10), and the HA antibody (sc7392) were purchased from Santa Cruz Biotechnology. The antibody against Flag (M2) was purchased from Sigma. The antibody against GAPDH (AC027) and p21 (A11877) were purchased from Abclonal. The antibody against Phospho-Histone H2A.X (Ser139) (2577 s) was purchased from CST.
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2

Immunohistochemical Analysis of Protein Expression

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Antigen retrieval was performed by autoclave treatment at 100℃ for 10 min using Target retrieval solution High P.H. (DAKO Cytomation, Kyoto, Japan). At rst, GST-pi polyclonal antibody (MBL, Nagoya, Japan), COX-2 polyclonal antibody (Cayman Chemical, Chicago, IL), P53 polyclonal antibody (FL393) (Santa Cruz, Santacruz, CA), and Smad4 monoclonal antibody (B-8) (Santa Cruz) was used. Proteins were identi ed with Envision kit (DAKO cytomation, Kyoto, Japan). They were colored by 3,3-Diaminobenzidine Tetrahydrochloride (DAB) (WAKO, Osaka, Japan), and their nuclei were stained by Gill's hematoxylin solution (WAKO). It was determined to be positive when more than 50% of the composing cells was stained.
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3

Immunohistochemical Analysis of Protein Expression

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Antigen retrieval was performed by autoclave treatment at 100℃ for 10 min using Target retrieval solution High P.H. (DAKO Cytomation, Kyoto, Japan). At rst, GST-pi polyclonal antibody (MBL, Nagoya, Japan), COX-2 polyclonal antibody (Cayman Chemical, Chicago, IL), P53 polyclonal antibody (FL393) (Santa Cruz, Santacruz, CA), and Smad4 monoclonal antibody (B-8) (Santa Cruz) was used. Proteins were identi ed with Envision kit (DAKO cytomation, Kyoto, Japan). They were colored by 3,3-Diaminobenzidine Tetrahydrochloride (DAB) (WAKO, Osaka, Japan), and their nuclei were stained by Gill's hematoxylin solution (WAKO). It was determined to be positive when more than 50% of the composing cells was stained.
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