Application

Research CategoryEpigenetics & Nuclear Function

Research Sub CategoryCell Cycle, DNA Replication & Repair

This Anti-RecQ5 Antibody is validated for use in Western Blotting for the detection of RecQ5.

Western Blotting Analysis: A representative lot detected FLAG-tagged wild-type RecQ5 and a D157A (DA) ATPase-defective RecQ5 mutant construct, but not a helicase domain-truncated (a.a. 450–991 deleted) RECQ5 construct expressed in HEK293 T cells devoid of endogenous RecQ5 (Li, M., et al. (2015). Nat. Commun. 6:6720).
Western Blotting Analysis: Representative lots detected siRNA-mediated RecQ5 downregulation using whole cell lysate or chromatin fraction from human HEK293T cells. Combined treatment with siRNA and the transcription inhibitor 5,6-dichloro-beta-d-ribofuranosyl benzimidazole (DRB) further decreased cellular RecQ5 protein level (Li, M., et al. (2015). Nat. Commun. 6:6720; Li, M., et al. (2011). Mol. Cell. Biol. 31(10) 2090-2099).

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

ATP-dependent DNA helicase Q5 (EC 3.6.4.12; UniProt O94762; also known as DNA helicase RecQ-like type 5, RecQ5, RecQ protein-like 5) is encoded by the RECQL5 (also known as RECQ5) gene (Gene ID 9400) in human. Human RECQ proteins (RECQ1, BLM, WRN, RECQ4, and RECQ5) constitute a family of highly conserved DNA helicases with nonredundant functions to ensure proper development. RECQ5 plays an important role in preventing both transcription-associated double-stranded breaks (DSBs) and DSBs caused by TOP1 poisons, such as camptothecin (CPT), and Recq5 deficiency leads to genome instability and cancer development in mice. RECQ5 KIX domain mediated interaction with RNA polymerase II (RNAPII) prevents RNAPII from binding transcription elongation factor TFIIS, thereby suppressing RNAPII-dependent transcription initiation and DSBs. On the other hand, RECQ5 is also known to interact with the active RNAPII (RNAPIIo) elongation complex to maintain the transcription elongation rate and prevent genome instability. In addition, RECQ5 helicase domain mediated interaction with TOP1 is reported to facilitate TOP1 K391/K436 SUMOylation, promoting TOP1-RNAPIIo association and efficient recruitment of mRNA processing factors to the transcriptionally active chromatin to prevent genome instability caused by R-loops formation.

Immunogen

Linear peptide corresponding a sequence from the RAD51 interaction domain of human RecQ5.

Epitope: RAD51 interaction domain.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Rabbit polyclonal serum with 0.05% sodium azide.

Unpurified

Quality

Evaluated by Western Blotting in Raji nuclear extract.

Western Blotting Analysis: An 1:500 dilution of this antibody detected RecQ5 in 10 µg of Raji nuclear extract.

Specificity

This antiserum recognizes RECQ5 isoform Beta by targeting a sequence from the RAD51-interacting region within the helicase domain. This antiserum is expected to react with also spliced isoform 4, but not the helicase domain-truncated Alpha (RecQ5b) and Gamma (RecQ5a) spliced variants reported by UniProt (O94762).

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

~120 kDa observed. 108.9 kDa (isoform Beta) and 106.1 kDa (isoform 4) calculated.