u-2 os cell line Search Results


u-2 os  (ATCC)
99
ATCC u-2 os
U 2 Os, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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u 2 os cells  (Genecopoeia)
95
Genecopoeia u 2 os cells
U 2 Os Cells, supplied by Genecopoeia, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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u2os cell lysate  (Santa Cruz Biotechnology)
91
Santa Cruz Biotechnology u2os cell lysate
PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the <t>U2OS</t> <t>cell</t> line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.
U2os Cell Lysate, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human bone osteosarcoma cell line u-2 os  (Human Protein Atlas)
90
Human Protein Atlas human bone osteosarcoma cell line u-2 os
PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the <t>U2OS</t> <t>cell</t> line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.
Human Bone Osteosarcoma Cell Line U 2 Os, supplied by Human Protein Atlas, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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u2os human osteosarcoma cell line  (Elabscience Biotechnology)
90
Elabscience Biotechnology u2os human osteosarcoma cell line
PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the <t>U2OS</t> <t>cell</t> line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.
U2os Human Osteosarcoma Cell Line, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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u-2 os  (European Collection of Authenticated Cell Cultures)
90
European Collection of Authenticated Cell Cultures u-2 os
PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the <t>U2OS</t> <t>cell</t> line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.
U 2 Os, supplied by European Collection of Authenticated Cell Cultures, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human bone cell line u-2-os  (MARINPHARM gmbh)
90
MARINPHARM gmbh human bone cell line u-2-os
PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the <t>U2OS</t> <t>cell</t> line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.
Human Bone Cell Line U 2 Os, supplied by MARINPHARM gmbh, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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bone osteosarcoma cell line u2-os  (Molecular Medicine LLC)
90
Molecular Medicine LLC bone osteosarcoma cell line u2-os
PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the <t>U2OS</t> <t>cell</t> line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.
Bone Osteosarcoma Cell Line U2 Os, supplied by Molecular Medicine LLC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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u-2 os cells  (Korean Cell Line Bank)
90
Korean Cell Line Bank u-2 os cells
PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the <t>U2OS</t> <t>cell</t> line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.
U 2 Os Cells, supplied by Korean Cell Line Bank, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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u-2 os ht-ecs cell line  (Promega)
90
Promega u-2 os ht-ecs cell line
PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the <t>U2OS</t> <t>cell</t> line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.
U 2 Os Ht Ecs Cell Line, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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u-2 os cell line  (AstraZeneca ltd)
90
AstraZeneca ltd u-2 os cell line
PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the <t>U2OS</t> <t>cell</t> line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.
U 2 Os Cell Line, supplied by AstraZeneca ltd, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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u2-os cells  (Korean Cell Line Bank)
90
Korean Cell Line Bank u2-os cells
PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the <t>U2OS</t> <t>cell</t> line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.
U2 Os Cells, supplied by Korean Cell Line Bank, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the U2OS cell line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.

Journal:

Article Title: General Transcriptional Coactivator PC4 Activates p53 Function

doi: 10.1128/MCB.24.5.2052-2062.2004

Figure Lengend Snippet: PC4 directly interacts with p53 in vitro and in vivo. (A) Induction of p53 expression in the U2OS cell line. The levels of p53 and PC4 present in adriamycin (2 μg/ml)-treated U2OS cells were assessed by Western blotting using anti-p53 (upper panel) and anti-PC4 (lower panel) antibodies. (B and C) In vivo interaction of PC4 with p53. (B) Lane 1, an adriamycin (2 μg/ml)-induced U2OS cell extract was immunoblotted with polyclonal PC4 antibody N17. Lane 2, immunoprecipitation of endogenous PC4 from an induced U2OS cell lysate was performed using anti-p53 monoclonal antibody DO1 followed by immunoblotting with anti-PC4 polyclonal antibody. Lane 3, immunoprecipitation using mouse preimmune serum used as a control. (C) Lane 1, immunoprecipitation of endogenous p53 from an induced U2OS cell extract, using anti-PC4 polyclonal antibody N17 followed by immunoblotting with anti-p53 monoclonal antibody DO1. Lane 2, adriamycin-induced U2OS cell extract immunoblotted with monoclonal p53 antibody DO1. Lane 3, immunoprecipitation reaction with goat preimmune serum used as a control. (D, E, and F) Interaction of PC4 with p53 in an in vitro GST pulldown assay. (D) Schematic representation of GST and GST-p53 fusion proteins. ++, strong interaction of PC4 with respective GST-p53 fusion protein; +, weaker interaction; −, no interaction. (E) SDS-PAGE (10%) and Coomassie blue R250 staining of immobilized GST-p53 fusion proteins. Lane 1, GST-p53 (full length); lane 2, GST-p53(1-73), lane 3, GST-p53(120-290); lane 4, GST-p53(284-330), lane 5, GST-p53(328-368); lane 6, GST-p53(364-393). GST fusion proteins predominantly contain intact proteins (indicated with asterisks) with minimum low-molecular-weight breakdown products. (F) One microgram of GST (lane 2) or GST-p53 fusion proteins (lanes 3 to 8) was incubated with bacterial extract containing 200 ng of PC4 and analyzed by immunoblotting with anti-PC4 N17 antibody. Lane 1, 5% input of bacterial cell lysate. IP, immunoprecipitation; IB, immunoblot; WCE, whole-cell extract; AD, activation domain; DBD, DNA binding domain; OD, oligomerization domain.

Article Snippet: Immunoprecipitation was performed by incubating the drug-treated U2OS cell lysate with protein A-agarose beads conjugated to either anti-p53 mouse monoclonal antibody DO1 (Oncogene) or anti-PC4 goat polyclonal antibody N17 (Santa Cruz).

Techniques: In Vitro, In Vivo, Expressing, Western Blot, Immunoprecipitation, GST Pulldown Assay, SDS Page, Staining, Molecular Weight, Incubation, Activation Assay, Binding Assay