Determining protein partners is an essential step toward understanding protein function and identifying relevant biological pathways. Many methods exist for investigating protein–protein interactions. The pull-down assay is an in vitro technique used to detect physical interactions between two or more prot...
Protein dialysis and other purification techniques Immunoprecipitation and pull-down assays Other methods for protein preparation Download the Protein Preparation Handbook Principles of diffusion Dialysis works by diffusion, a process that result...
(cobalt) agarose beads. The fusion-tagged protein acts as the bait to capture a putative binding partner (i.e., the prey). In a typical pull-down assay, the immobilized bait protein is incubated with a cell lysate. After the prescribed washing steps, the 'interactors are selectively eluted...
Pull-down assay demonstrating in vitro protein-protein interaction between BmCRT and HuC1q. (A) Visualization of BmCRT-HuC1Q complex formation on 12% SDS-PAGE.Sunita, Yadav
We have previously identified some paraspeckle proteins as putative poly-PR-binding proteins (TableS1)10. In the current study, using GST pull-down assay with GST-FLAG-tagged PR100 as a bait, we confirmed the interaction between poly-PR and endogenous or exogenous paraspeckle proteins, hnRNPF,...
Purified protein pull-down assay The pull-down was performed as previously described60. 3 μg of recombinant Myc-PPP6R3 protein (TP328606, OriGene), GST-PPP6C protein (H00005537-P01, Novus), His-pAMPK (P48-10H-10, SignalChem) or His-AMPK (P48-14H-20, SignalChem) were mixed. The...
DPC pull-down The DPC pull-down assay is a high-stringency plasmid pull-down protocol to specifically isolate DNA crosslinked HpaII from extract (Larsen et al., 2018). Briefly, Streptavidin-coupled magnetic beads (Dynabeads M-280, Invitrogen; 10 μL per pull-down) were washed twice with Was...
UponreceiptstorePull-DownLysisBufferatroomtemperature.Storeremainingkitcomponentsat4°C.Kitisshippedatambienttemperature.TableofContentsIntroduction...2ImportantProductInformation...
Features of His Tag Protein Interaction Pull-Down Kit:• 6xHis pull-down (Product No. 21277)—purifies protein interactors of any His-tagged fusion protein• Complete kits—provide all components and detailed protocol for purifying protein:protein interactions• No special equipment needed—use ...
To further confirm that this binding sequence in the GPER promoter is ZNF32 specific, a DNA-oligo pull-down assay was carried out in ZR-75-30 cells with a specifically synthesized oligonucleotide containing the ZNF32-binding sequence (−1623 bp GCATTT) of the GPER gene and a synthesized ...