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rabbit polyclonal antibody against pp2ac  (Bio-Rad)
96
Bio-Rad rabbit polyclonal antibody against pp2ac
Rabbit Polyclonal Antibody Against Pp2ac, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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western immunoblots ephx1  (Gentest Corp)
90
Gentest Corp western immunoblots ephx1
Western Immunoblots Ephx1, supplied by Gentest Corp, 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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c fos antibody  (Santa Cruz Biotechnology)
96
Santa Cruz Biotechnology c fos antibody
Figure 1. Binding of SUMOylated c-Fos to a TRE containing promoter. (a) Immunoblotting characterization of the anti-SUMOylated c-Fos antibody. GST fusions of either c-Fos or c-Fos K265R were subjected to in vitro SUMOylation (ATP was used to start the reaction) as <t>previously</t> <t>described7</t> and analyzed by immunoblotting using a c-Fos antibody <t>(sc52,</t> Santa Cruz Biotechnologies, Santa Cruz, CA, USA, left panel) or the purified c-Fos-SUMO antibody (see Supplementary Figure 1 for details on its generation, right panel). The arrow indicates SUMOylated c-Fos. (b, c) Binding of SUMOylated c-Fos to the 4 TRE promoter. Human HA-tagged c-Fos and c-Fos-K265R were transferred into the pCDNA5/FRT/ TO/SH/GW plasmid30 using the Gateway technology (Invitrogen, Carlsbad, CA, USA) and stably inserted in Flip-In T-Rex 293 cells at their unique FRT site by recombination following to the manufacturer’s protocol (Invitrogen). These cells were also stably transfected with a 4 TRE-luc reporter gene17 and relative quantitative PCR (qPCR) quantification showed that two copies of the transgene were integrated in their genome (data not shown). Cells were seeded (5.106 cells per 10 cm diameter plate) and stimulated 72 h later with doxycycline (Dox; 100 ng/ml) and TPA (20 nM) for the indicated times before immunoblotting analysis (b) with anti-HA and -c-Fos-SUMO antibodies. (c) ChiPs were carried out after 2 h of induction with both antibodies. A pre-immune serum (immunoglobulin G (IgG)) was used as a nonspecific control in ChIPs. ChIPs were performed as previously described31 except that cells were fixed in 1% methanol-free paraformaldehyde at room temperature for 8 min. Nuclear lysates were sonicated using the Bioruptor UCD-200 sonifier (Diagenode, Liege, Belgium; 20 cycles of 30-s on and 30-s off, highest power). The data obtained using the Roche LightCycler 480 real-time qPCR device (Roche, Basel, Switzerland) were normalized to inputs taken from samples before immunoprecipitation and processed in parallel (n ¼ 3, means±s.d., **Po0.01).
C Fos Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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construct p1-6x-hcpro  (HCPro Inc)
90
HCPro Inc construct p1-6x-hcpro
Figure 1. Binding of SUMOylated c-Fos to a TRE containing promoter. (a) Immunoblotting characterization of the anti-SUMOylated c-Fos antibody. GST fusions of either c-Fos or c-Fos K265R were subjected to in vitro SUMOylation (ATP was used to start the reaction) as <t>previously</t> <t>described7</t> and analyzed by immunoblotting using a c-Fos antibody <t>(sc52,</t> Santa Cruz Biotechnologies, Santa Cruz, CA, USA, left panel) or the purified c-Fos-SUMO antibody (see Supplementary Figure 1 for details on its generation, right panel). The arrow indicates SUMOylated c-Fos. (b, c) Binding of SUMOylated c-Fos to the 4 TRE promoter. Human HA-tagged c-Fos and c-Fos-K265R were transferred into the pCDNA5/FRT/ TO/SH/GW plasmid30 using the Gateway technology (Invitrogen, Carlsbad, CA, USA) and stably inserted in Flip-In T-Rex 293 cells at their unique FRT site by recombination following to the manufacturer’s protocol (Invitrogen). These cells were also stably transfected with a 4 TRE-luc reporter gene17 and relative quantitative PCR (qPCR) quantification showed that two copies of the transgene were integrated in their genome (data not shown). Cells were seeded (5.106 cells per 10 cm diameter plate) and stimulated 72 h later with doxycycline (Dox; 100 ng/ml) and TPA (20 nM) for the indicated times before immunoblotting analysis (b) with anti-HA and -c-Fos-SUMO antibodies. (c) ChiPs were carried out after 2 h of induction with both antibodies. A pre-immune serum (immunoglobulin G (IgG)) was used as a nonspecific control in ChIPs. ChIPs were performed as previously described31 except that cells were fixed in 1% methanol-free paraformaldehyde at room temperature for 8 min. Nuclear lysates were sonicated using the Bioruptor UCD-200 sonifier (Diagenode, Liege, Belgium; 20 cycles of 30-s on and 30-s off, highest power). The data obtained using the Roche LightCycler 480 real-time qPCR device (Roche, Basel, Switzerland) were normalized to inputs taken from samples before immunoprecipitation and processed in parallel (n ¼ 3, means±s.d., **Po0.01).
Construct P1 6x Hcpro, supplied by HCPro Inc, 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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western blot  (SAS institute)
90
SAS institute western blot
Figure 1. Binding of SUMOylated c-Fos to a TRE containing promoter. (a) Immunoblotting characterization of the anti-SUMOylated c-Fos antibody. GST fusions of either c-Fos or c-Fos K265R were subjected to in vitro SUMOylation (ATP was used to start the reaction) as <t>previously</t> <t>described7</t> and analyzed by immunoblotting using a c-Fos antibody <t>(sc52,</t> Santa Cruz Biotechnologies, Santa Cruz, CA, USA, left panel) or the purified c-Fos-SUMO antibody (see Supplementary Figure 1 for details on its generation, right panel). The arrow indicates SUMOylated c-Fos. (b, c) Binding of SUMOylated c-Fos to the 4 TRE promoter. Human HA-tagged c-Fos and c-Fos-K265R were transferred into the pCDNA5/FRT/ TO/SH/GW plasmid30 using the Gateway technology (Invitrogen, Carlsbad, CA, USA) and stably inserted in Flip-In T-Rex 293 cells at their unique FRT site by recombination following to the manufacturer’s protocol (Invitrogen). These cells were also stably transfected with a 4 TRE-luc reporter gene17 and relative quantitative PCR (qPCR) quantification showed that two copies of the transgene were integrated in their genome (data not shown). Cells were seeded (5.106 cells per 10 cm diameter plate) and stimulated 72 h later with doxycycline (Dox; 100 ng/ml) and TPA (20 nM) for the indicated times before immunoblotting analysis (b) with anti-HA and -c-Fos-SUMO antibodies. (c) ChiPs were carried out after 2 h of induction with both antibodies. A pre-immune serum (immunoglobulin G (IgG)) was used as a nonspecific control in ChIPs. ChIPs were performed as previously described31 except that cells were fixed in 1% methanol-free paraformaldehyde at room temperature for 8 min. Nuclear lysates were sonicated using the Bioruptor UCD-200 sonifier (Diagenode, Liege, Belgium; 20 cycles of 30-s on and 30-s off, highest power). The data obtained using the Roche LightCycler 480 real-time qPCR device (Roche, Basel, Switzerland) were normalized to inputs taken from samples before immunoprecipitation and processed in parallel (n ¼ 3, means±s.d., **Po0.01).
Western Blot, supplied by SAS institute, 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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c terminal region  (Danaher Inc)
99
Danaher Inc c terminal region
Figure 1. Binding of SUMOylated c-Fos to a TRE containing promoter. (a) Immunoblotting characterization of the anti-SUMOylated c-Fos antibody. GST fusions of either c-Fos or c-Fos K265R were subjected to in vitro SUMOylation (ATP was used to start the reaction) as <t>previously</t> <t>described7</t> and analyzed by immunoblotting using a c-Fos antibody <t>(sc52,</t> Santa Cruz Biotechnologies, Santa Cruz, CA, USA, left panel) or the purified c-Fos-SUMO antibody (see Supplementary Figure 1 for details on its generation, right panel). The arrow indicates SUMOylated c-Fos. (b, c) Binding of SUMOylated c-Fos to the 4 TRE promoter. Human HA-tagged c-Fos and c-Fos-K265R were transferred into the pCDNA5/FRT/ TO/SH/GW plasmid30 using the Gateway technology (Invitrogen, Carlsbad, CA, USA) and stably inserted in Flip-In T-Rex 293 cells at their unique FRT site by recombination following to the manufacturer’s protocol (Invitrogen). These cells were also stably transfected with a 4 TRE-luc reporter gene17 and relative quantitative PCR (qPCR) quantification showed that two copies of the transgene were integrated in their genome (data not shown). Cells were seeded (5.106 cells per 10 cm diameter plate) and stimulated 72 h later with doxycycline (Dox; 100 ng/ml) and TPA (20 nM) for the indicated times before immunoblotting analysis (b) with anti-HA and -c-Fos-SUMO antibodies. (c) ChiPs were carried out after 2 h of induction with both antibodies. A pre-immune serum (immunoglobulin G (IgG)) was used as a nonspecific control in ChIPs. ChIPs were performed as previously described31 except that cells were fixed in 1% methanol-free paraformaldehyde at room temperature for 8 min. Nuclear lysates were sonicated using the Bioruptor UCD-200 sonifier (Diagenode, Liege, Belgium; 20 cycles of 30-s on and 30-s off, highest power). The data obtained using the Roche LightCycler 480 real-time qPCR device (Roche, Basel, Switzerland) were normalized to inputs taken from samples before immunoprecipitation and processed in parallel (n ¼ 3, means±s.d., **Po0.01).
C Terminal Region, supplied by Danaher Inc, 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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Image Search Results


Figure 1. Binding of SUMOylated c-Fos to a TRE containing promoter. (a) Immunoblotting characterization of the anti-SUMOylated c-Fos antibody. GST fusions of either c-Fos or c-Fos K265R were subjected to in vitro SUMOylation (ATP was used to start the reaction) as previously described7 and analyzed by immunoblotting using a c-Fos antibody (sc52, Santa Cruz Biotechnologies, Santa Cruz, CA, USA, left panel) or the purified c-Fos-SUMO antibody (see Supplementary Figure 1 for details on its generation, right panel). The arrow indicates SUMOylated c-Fos. (b, c) Binding of SUMOylated c-Fos to the 4 TRE promoter. Human HA-tagged c-Fos and c-Fos-K265R were transferred into the pCDNA5/FRT/ TO/SH/GW plasmid30 using the Gateway technology (Invitrogen, Carlsbad, CA, USA) and stably inserted in Flip-In T-Rex 293 cells at their unique FRT site by recombination following to the manufacturer’s protocol (Invitrogen). These cells were also stably transfected with a 4 TRE-luc reporter gene17 and relative quantitative PCR (qPCR) quantification showed that two copies of the transgene were integrated in their genome (data not shown). Cells were seeded (5.106 cells per 10 cm diameter plate) and stimulated 72 h later with doxycycline (Dox; 100 ng/ml) and TPA (20 nM) for the indicated times before immunoblotting analysis (b) with anti-HA and -c-Fos-SUMO antibodies. (c) ChiPs were carried out after 2 h of induction with both antibodies. A pre-immune serum (immunoglobulin G (IgG)) was used as a nonspecific control in ChIPs. ChIPs were performed as previously described31 except that cells were fixed in 1% methanol-free paraformaldehyde at room temperature for 8 min. Nuclear lysates were sonicated using the Bioruptor UCD-200 sonifier (Diagenode, Liege, Belgium; 20 cycles of 30-s on and 30-s off, highest power). The data obtained using the Roche LightCycler 480 real-time qPCR device (Roche, Basel, Switzerland) were normalized to inputs taken from samples before immunoprecipitation and processed in parallel (n ¼ 3, means±s.d., **Po0.01).

Journal: Oncogene

Article Title: SUMOylation of the inducible (c-Fos:c-Jun)/AP-1 transcription complex occurs on target promoters to limit transcriptional activation.

doi: 10.1038/onc.2013.4

Figure Lengend Snippet: Figure 1. Binding of SUMOylated c-Fos to a TRE containing promoter. (a) Immunoblotting characterization of the anti-SUMOylated c-Fos antibody. GST fusions of either c-Fos or c-Fos K265R were subjected to in vitro SUMOylation (ATP was used to start the reaction) as previously described7 and analyzed by immunoblotting using a c-Fos antibody (sc52, Santa Cruz Biotechnologies, Santa Cruz, CA, USA, left panel) or the purified c-Fos-SUMO antibody (see Supplementary Figure 1 for details on its generation, right panel). The arrow indicates SUMOylated c-Fos. (b, c) Binding of SUMOylated c-Fos to the 4 TRE promoter. Human HA-tagged c-Fos and c-Fos-K265R were transferred into the pCDNA5/FRT/ TO/SH/GW plasmid30 using the Gateway technology (Invitrogen, Carlsbad, CA, USA) and stably inserted in Flip-In T-Rex 293 cells at their unique FRT site by recombination following to the manufacturer’s protocol (Invitrogen). These cells were also stably transfected with a 4 TRE-luc reporter gene17 and relative quantitative PCR (qPCR) quantification showed that two copies of the transgene were integrated in their genome (data not shown). Cells were seeded (5.106 cells per 10 cm diameter plate) and stimulated 72 h later with doxycycline (Dox; 100 ng/ml) and TPA (20 nM) for the indicated times before immunoblotting analysis (b) with anti-HA and -c-Fos-SUMO antibodies. (c) ChiPs were carried out after 2 h of induction with both antibodies. A pre-immune serum (immunoglobulin G (IgG)) was used as a nonspecific control in ChIPs. ChIPs were performed as previously described31 except that cells were fixed in 1% methanol-free paraformaldehyde at room temperature for 8 min. Nuclear lysates were sonicated using the Bioruptor UCD-200 sonifier (Diagenode, Liege, Belgium; 20 cycles of 30-s on and 30-s off, highest power). The data obtained using the Roche LightCycler 480 real-time qPCR device (Roche, Basel, Switzerland) were normalized to inputs taken from samples before immunoprecipitation and processed in parallel (n ¼ 3, means±s.d., **Po0.01).

Article Snippet: GST fusions of either c-Fos or c-Fos K265R were subjected to in vitro SUMOylation (ATP was used to start the reaction) as previously described7 and analyzed by immunoblotting using a c-Fos antibody (sc52, Santa Cruz Biotechnologies, Santa Cruz, CA, USA, left panel) or the purified c-Fos-SUMO antibody (see Supplementary Figure 1 for details on its generation, right panel).

Techniques: Binding Assay, Western Blot, In Vitro, Stable Transfection, Transfection, Real-time Polymerase Chain Reaction, Control, Sonication, Immunoprecipitation