Recombinant Human RAD51, GST-tagged

• Cat.No.: RAD51-2155H
• Product Overview: Recombinant Human RAD51 protein, fused to GST-tag, was expressed in E.coli and purified by GSH-sepharose.

Specification

• Species: Human
• Source: E.coli
• Tag: GST
• Protein Length: 1-242aa
• Storage: The protein is stored in PBS buffer at -20℃. Avoid repeated freezing and thawing cycles.
• Storage Buffer: 1M PBS (58mM Na2HPO4,17mM NaH2PO4, 68mM NaCl, pH8. ) added with 100mM GSH and 1% Triton X-100,15%glycerol.

Gene Information

• Gene Name: RAD51 RAD51 homolog (S. cerevisiae) [ Homo sapiens ]
• Official Symbol: RAD51
• Synonyms: RAD51; RAD51 homolog (S. cerevisiae); RAD51 (S. cerevisiae) homolog (E coli RecA homolog) , RAD51 homolog (RecA homolog, E. coli) (S. cerevisiae) , RAD51A, RECA; DNA repair protein RAD51 homolog 1; BRCA1/BRCA2 containing complex; subunit 5; BRCC5; HsRad5
• Gene ID: 5888
• mRNA Refseq: NM_001164269
• Protein Refseq: NP_001157741
• MIM: 179617
• UniProt ID: Q06609
• Chromosome Location: 15q15.1
• Pathway: Assembly of the RAD51-ssDNA nucleoprotein complex, organism-specific biosystem; BARD1 signaling events, organism-specific biosystem; DNA Repair, organism-specific biosystem; Double-Strand Break Repair, organism-specific biosystem; Fanconi anemia pathway,
• Function: ATP binding; ATP binding; DNA-dependent ATPase activity; damaged DNA binding; double-stranded DNA binding; double-stranded DNA binding; identical protein binding; nucleoside-triphosphatase activity; nucleotide binding; protein C-terminus binding; protein

Citation

Synthetic Lethality in Pancreatic Cancer: Discovery of a New RAD51-BRCA2 Small Molecule Disruptor That Inhibits Homologous Recombination and Synergizes with Olaparib

Journal: Journal of Medicinal Chemistry    PubMed ID: 32037829    Data: 2020/2/10

Authors: Greta Bagnolini, Domenico Milano, Andrea Cavalli

Article Snippet:Competitive ELISA screening assay using biotinylated BRC4 peptide to disrupt the BRC4–RAD51 interaction was performed by modifying the method described by Rajendra et al. BRC4-biotinylated peptide (N-term biotin-KEPTLLGFHTASGKKVKIAKESLDKVKNLFDEKEQ from Life Technologies) was used to coat 384-well plates (Nunc).Competitive ELISA screening assay using biotinylated BRC4 peptide to disrupt the BRC4–RAD51 interaction was performed by modifying the method described by Rajendra et al. BRC4-biotinylated peptide (N-term biotin-KEPTLLGFHTASGKKVKIAKESLDKVKNLFDEKEQ from Life Technologies) was used to coat 384-well plates (Nunc).. After washing with PBS containing 0.05% Tween-20 (PBST) and blocking with the solution BSA 1% /PBST, overnight hybridization with human RAD51 protein (NP_002866 Creative BioMart, NY) was performed.. Test compounds were added in dose response from 0.01 to 100 μM in triplicate with constant DMSO 1%.Test compounds were added in dose response from 0.01 to 100 μM in triplicate with constant DMSO 1%.

(A) RAD51-BRCA2 BRC repeat complex (PDB code 1N0W). RAD51 is represented as a surface, BRC4 as a cartoon. The two hot spots of the interaction between the proteins (Phe1524 and Phe1546) are highlighted in sticks. (B) Zone II magnification showing the interacting residues of BRC4 (yellow) and RAD51 (white).

Both enantiomers of compound 4d docked into the LFDE binding site (site II) of RAD51 (PDB code 1N0W).

MST analysis of His-hRAD51- 35d binding. Titration curve of (RED-tris-NTA 2nd Generation)-His-hRAD51 (80 nM) with increasing concentrations of 35d . Sigmoidal fitting curve was obtained using the Affinity Analysis software of NanoTemper Technologies. MST data are the average of three replicates.

Rad51/BRCA2 disruptors inhibit homologous recombination and synergize with olaparib in pancreatic cancer cells.

Journal: European journal of medicinal chemistry    PubMed ID: 30660828    Data: 2019/3/21

Authors: Marinella Roberti, Fabrizio Schipani, Andrea Cavalli

Article Snippet:M AN US CR IP T AC CE PT ED 31 BRC4-biotinylated peptide (N-term BiotinKEPTLLGFHTASGKKVKIAKESLDKVKNLFDEKEQ from Life Technologies) was used to coat 384-well plates (Nunc).M AN US CR IP T AC CE PT ED 31 BRC4-biotinylated peptide (N-term BiotinKEPTLLGFHTASGKKVKIAKESLDKVKNLFDEKEQ from Life Technologies) was used to coat 384-well plates (Nunc).. After washing with PBS containing 0.05% Tween-20 (PBST), and blocking with the solution BSA 1% /PBST, overnight hybridization with human Rad51 protein (NP_002866 Creative BioMart, NY) was performed.. Test compounds were added in dose-response from 0.01 to 100μM in triplicate with constant DMSO 1%.Test compounds were added in dose-response from 0.01 to 100μM in triplicate with constant DMSO 1%.