Anti-HPV16E6 antibodies 1E-6F4 (E6 N-ter) and 2E-3F8 (E6 C-ter) were obtained from Euromedex. Anti-p53 (DO-1) and anti-pRb (4H1) antibodies were obtained from Cell Signaling. Anti-β-actin (AC-15) antibody was obtained from Sigma-Aldrich. Anti-HPV16E7 (NM2) antibody was obtained from Santa Cruz Biotechnology. HRP-conjugated goat anti-mouse secondary antibody was purchased from BD Pharmingen. Primary and secondary antibodies have been diluted respectively to 1:1000 and 1:5000 in blocking buffer (except for β-actin at 1:20,000 and 1:30,000).
Anti p53 do 1
Manufactured by Cell Signaling Technology
Sourced in China, United States
Anti-p53 (DO-1) is a mouse monoclonal antibody that recognizes the human p53 protein. It binds to the N-terminal region of p53 and can be used for the detection of p53 in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin antibody ac 15, by Merck Group (1 mentions)
Anti prb 4h1, by Cell Signaling Technology (1 mentions)
Anti cbp, by Cell Signaling Technology (1 mentions)
Anti puma, by Santa Cruz Biotechnology (1 mentions)
Anti p21, by Santa Cruz Biotechnology (1 mentions)
Anti mdm2, by Santa Cruz Biotechnology (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Anti p ser1981 atm, by Abcam (1 mentions)
Irdye 800cw goat anti mouse, by LI COR (1 mentions)
Irdye 800cw goat anti rabbit, by LI COR (1 mentions)
4 protocols using anti p53 do 1
1
Immunodetection of HPV16 Proteins
2
Western Blot Assays with Antibodies and Plasmids
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The following antibodies were used in western blot assays: anti-β-actin (A15), anti-Flag M2 and anti-Flag M2 agarose resin (Sigma), anti-HA (3F10), anti-HA agarose resin (Roche Applied Science), anti-p53 (DO-1, which detects the N-terminal epitope comprising amino acids 11–25), anti-p53 (full-length), anti-CBP(Cell Signaling Technology), anti-p21, anti-PUMA, anti-MDM2 (Santa Cruz), anti-Ac-K101-p53, anti-Ac-K164-p53, anti-Ac-p53 C-terminal (made in-house), anti-pan-acetylation-Lysine (Ac-K) and anti-pan β-hydroxybutyrylation-Lysine (BHB-K) (PTM BioLabs; Hangzhou, China).
Plasmids Flag-p53, Flag-p300, CBP-HA and CBP-HA mutants were from Dr. Gu’s lab4 (link),8 (link).
siRNA targeting CBP and p300 (1#: 5′-GAGGUCGUUUACAUAAATT-3′; 2#: 5′-UUUAUGUAAACGCGACCUCTT-3′) and a negative control (5′-UUCUCCGAACGUGUCACGUTT-3′) were synthesized by GenePharma (China).
Plasmids Flag-p53, Flag-p300, CBP-HA and CBP-HA mutants were from Dr. Gu’s lab4 (link),8 (link).
siRNA targeting CBP and p300 (1#: 5′-GAGGUCGUUUACAUAAATT-3′; 2#: 5′-UUUAUGUAAACGCGACCUCTT-3′) and a negative control (5′-UUCUCCGAACGUGUCACGUTT-3′) were synthesized by GenePharma (China).
3
Western Blot Analysis of DNA Damage Response
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Western blot and detection were performed as previously reported (36 (link)). The activation state of DNA damage response proteins was analyzed by using the following antidodies: rabbit mAb anti-Ser1981 p-ATM (Abcam Ltd.); mouse mAb anti-p53 DO-1 and rabbit pAb anti-Thr68 p-CHK2 (Cell Signaling, Beverly, MA, USA). As loading control, levels of β-ACTIN were evaluated by using the mouse mAb anti-β-actin (Sigma-Aldrich).
4
Plasmid Construction and Lentivirus Generation
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Expression plasmids of pRK-Flag-hCINAP and pRK-HA-RPS14 were constructed by inserting full-length complementary DNAs of hCINAP or RPS14 into the pRK vector at EcoRI and XbaI restriction sites. Insertions were verified by DNA sequencing. To generate recombinant lentivirus vectors expressing hCINAP-shRNA or non-silencing control-shRNA hCINAP-shRNA-1: 5′- CAGAGUAGUUGAUGAGUUA -3′, hCINAP-shRNA-2: 5′- GAGAGAAGGUGGAGUUAUU -3′ and non-silencing control-shRNA: 5′- UUCUCCGAACGUGUCACGU -3′ were cloned into the pGCSIL-Puromycin lentivirus vector (GeneChem Co. Ltd, China). Antibodies including anti-Myc (M047-3), anti-GST (M071-3), anti-β-actin (PM053), anti-Lamin B1 (PM064), anti-His (D291-3) and anti-α-Tubulin (M175-3) were purchased from MBL. Monoclonal anti-Flag (M2; F3165) and anti-HA(HA-7; H9658) were from Sigma. Anti-p53 (DO-1) was from Cell Signaling Technology. IRDye 800CW Goat anti-mouse (926–32210) and IRDye 800CW Goat anti-rabbit (926–32211) were purchased from LI-COR Bioscience. Fluorescein isothiocyanate-conjugated goat anti-mouse IgG (ZF-0312) and TRITC-conjugated anti-rabbit IgG (ZF-0316) were from ZSGB-Bio. Rabbit polyclonal anti-hCINAP was generated as previously described25 (link).
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