Journal: STAR Protocols

Article Title: Protocol for the genome-wide identification of intrinsic transcription factor binding motifs by mammalian-optimized pull-down sequencing

doi: 10.1016/j.xpro.2026.104513

Figure Lengend Snippet: Expression and purification of recombinant His-MBP-FOXP3(ΔN) protein (A) Schematic flowchart of the protein purification process, including Ni-NTA affinity chromatography followed by HiTrap Heparin HP chromatography. (B) SDS-PAGE analysis of purified fractions during the HiTrap Heparin HP chromatography. The arrow indicates the target protein. M represents the molecular weight marker, with numbers on the right indicating molecular weights in kDa. The gel was stained with Coomassie Blue dye. (C) The HiTrap Heparin HP affinity chromatography profile of the final purified protein. An asterisk indicates the peak corresponding to the desired FOXP3.

Article Snippet: Alternatives: This protocol uses NEBNext Ultra II DNA Library Prep Kit with Sample Purification Beads (NEB #E7103S) to construct library.

Techniques: Expressing, Purification, Recombinant, Protein Purification, Affinity Chromatography, Chromatography, SDS Page, Molecular Weight, Marker, Staining

Journal: STAR Protocols

Article Title: Protocol for the genome-wide identification of intrinsic transcription factor binding motifs by mammalian-optimized pull-down sequencing

doi: 10.1016/j.xpro.2026.104513

Figure Lengend Snippet: Library preparation Schematic diagram illustrating the major steps of library construction using the NEBNext Ultra II DNA Library Prep Kit. The workflow includes. (1) end repair and dA-tailing of fragmented genomic DNA. (2) adaptor ligation (hairpin adaptor containing dU); USER enzyme cleavage to open the adaptor, and (3) PCR enrichment with indexed primers to generate the final sequencing library.

Article Snippet: NEBNext Ultra II DNA Library Prep with Sample Purification Beads , NEB , Cat#E7103S.

Techniques: Ligation, Sequencing

Journal: STAR Protocols

Article Title: Protocol for the genome-wide identification of intrinsic transcription factor binding motifs by mammalian-optimized pull-down sequencing

doi: 10.1016/j.xpro.2026.104513

Figure Lengend Snippet: Library preparation Schematic diagram illustrating the major steps of library construction using the NEBNext Ultra II DNA Library Prep Kit. The workflow includes. (1) end repair and dA-tailing of fragmented genomic DNA. (2) adaptor ligation (hairpin adaptor containing dU); USER enzyme cleavage to open the adaptor, and (3) PCR enrichment with indexed primers to generate the final sequencing library.

Article Snippet: Alternatives: This protocol uses NEBNext Ultra II DNA Library Prep Kit with Sample Purification Beads (NEB #E7103S) to construct library.

Techniques: Ligation, Sequencing