Immunoprecipitation of endogenous proteins was accomplished using a Universal Co-IP kit (Cat. No. 54002, Active Motif). HCT116 or HT29 nuclear extracts were first cleared with protein A agarose beads, then incubated with HIF-2α antibody (Cat. No. NB100-132, Novus Biologicals, Littleton, CO) or normal mouse IgG (Cat. No. sc-2025, Santa Cruz Biotechnology, Santa Cruz, CA) for 2 hr before addition of protein A agarose beads. After binding, beads were pelleted by centrifugation and washed with buffer, then immunoprecipitated materials were eluted and immunoblotted with anti-human p300 (1:500 dilution; Cat. No. sc-584, Santa Cruz Biotechnology, Santa Cruz, CA), anti-human CBP (1:500 dilution; Cat. No. 4772, Cell Signaling Technology, Danvers, MA), or anti-HIF-2α (1:1,000 dilution; Cat. No. NB100-132, Novus Biologicals) primary antibodies.
Anti human cbp
Manufactured by Santa Cruz Biotechnology
Sourced in Canada
Anti-human CBP is a laboratory reagent designed for use in various research applications. It specifically targets the CREB-binding protein (CBP), a transcriptional coactivator that plays a crucial role in gene regulation. This product can be utilized in techniques such as immunoprecipitation, Western blotting, and other assays to study the interactions and functions of CBP within cellular systems.
Automatically generated - may contain errors
4 protocols using anti human cbp
1
Immunoprecipitation of Endogenous Proteins
2
Immunoprecipitation of EPAS1 Interactors
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Immunoprecipitation of endogenous proteins was accomplished with a Universal Co-IP kit (Cat. No. 54002, Active Motif) according to the manufacturer’s protocol. HT1080 nuclear extracts were first incubated with protein A agarose beads. Cleared supernatants were incubated with EPAS1 (HIF-2α) antibody (Cat. No. NB100-132, Novus Biologicals) or normal mouse IgG (Cat. No. sc-2025, Santa Cruz Biotechnology, Santa Cruz, CA) 2 hr before addition of protein A agarose beads. After binding, beads were pelleted by centrifugation and washed. After washing, immunoprecipitated materials were eluted and immunoblotted with anti-human p300 (1:500 dilution; Cat. No. sc-584, Santa Cruz Biotechnology), anti-human CBP (1:500 dilution; Cat. No. 4772, Cell Signaling Technology), or anti-human HIF-2α mouse monoclonal (1:1,000 dilution; Cat. No. NB100-132, Novus Biologicals) antibodies.
3
Endogenous Protein Co-Immunoprecipitation
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Immunoprecipitation of endogenous proteins was accomplished using a Universal Co-IP kit (Cat. No. 54002, Active Motif). HT1080 nuclear extracts were first incubated with protein A agarose beads. Cleared supernatants were then incubated with HIF-2α antibody (Cat. No. NB100-132, Novus Biologicals) or normal mouse IgG (Cat. No. sc-2025, Santa Cruz Biotechnology) for 2 hr before addition of protein A agarose beads. After binding, beads were pelleted by centrifugation and washed with buffer. After washing, immunoprecipitated materials were eluted and immunoblotted with anti-human p300 (1:500 dilution; Cat. No. sc-584, Santa Cruz Biotechnology), anti-human CBP (1:500 dilution; Cat. No. 4772, Cell Signaling Technology), or anti-HIF-2α (1:1,000 dilution; Cat. No. NB100-132, Novus Biologicals) primary antibodies.
4
Protein Extraction and Immunoblotting Assay
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To prepare extracts, we used CytoBuster protein extraction reagent (Cat. No. 71009, Novagen, Gibbstown, NJ) or NE-PER nuclear and cytoplasmic extraction reagents (Cat. No. 78833, Pierce, Rockford, IL) as previously described [9 (link)]. Proteins were analyzed by immunoblotting with anti-human p300 (1:500 dilution; Cat. No. sc-584, Santa Cruz Biotechnology), anti-human CBP (1:500 dilution; Cat. No. 4772, Cell Signaling Technology), anti-human ACLY (1:1,000 dilution; Cat. No. 4332, Cell Signaling Technology), anti-human ACSS1 (1:1,000 dilution; Cat. No. SAB1400745, Sigma), anti-human ACSS2 (1:500 dilution; Cat. No. ab66038, Abcam, Cambridge, MA), anti-human HIF-2α (1:1,000 dilution; Cat. No. NB100-132, Novus Biologicals), anti-TATA-binding protein (TBP) (1:1,000 dilution; Cat. No. sc-204, Santa Cruz Biotechnology), anti-α-tubulin (1:10,000 dilution; Cat. No. T9026, Sigma), or anti-HA (1:5,000 dilution; Cat. No. H9658, Sigma) antibodies as indicated.
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