Description
General description
GTPase HflX (UniProt: O33230; also known as GTP-binding protein HflX) is encoded by the hflX (also known as Rv2725c) gene (Gene ID: 888241) in Mycobacterium tuberculosis. High frequency of lysogenization X (HflX) protein belongs to the superfamily of the Obg-HflX like ucpGTPases class of translation factors that is highly conserved in the entire prokaryotic kingdom. It is involved in the stress response of bacteria and participates in protein translation, likely by providing energy for protein synthesis, or by facilitating the recycling of factors involved in translation. HflX is a ribosome-binding protein that binds to the E-site of 70S bacterial ribosomes and induces split into 50S/30S ribosomal subunits upon GTP hydrolysis. HflX produced by slow-growing Mycobacterium bovis (M. bovis) is reported to play a critical role in the pathogen s transition to a nonreplicating, drug-tolerant state in response to hypoxia. On the other hand, HflX-deficient M. bovis (knockout) replicate at a markedly faster rate in the microaerophilic phase of a hypoxia model, which results in premature entry into dormancy. The absence of HflX also results in a significant accumulation of 70S ribosomes upon erythromycin exposure, and deletion of its N-terminal or the C-terminal domain abrogates ribosome splitting and abolishes its ability to mediate antibiotic tolerance. HflX is overexpressed in M. bovis exposed to the hostile lysosomal environment of macrophages. Under normoxia culture conditions it does not play any essential role in M. bovis, but under hypoxic state it modulates translational activity and controls Mycobacterium growth rate. (Ref.: Ngan, JYG., et al. (2021) Proc. Natl. Acad. Sci. USA. 118(12); e2006717118; Rudra, P., et al., (2020). Proc. Natl. Acad. Sci. USA. 117(1); 629-634).)
Immunogen
His-tagged recombinant GTPase HflX from Mycobacterium tuberculosis.
Application
Quality Control Testing
Evaluated by Dot Blot Analysis with His-tagged, recombinant GTPase HfLX from Mycobacterium tuberculosis.
Dot Blot Analysis: A 1:1,000 dilution of this antibody detected His-tagged, recombinant GTPase HflX from mycobacterium tuberculosis.
Tested Applications
ELISA Analysis: A representative lot detected HfIX in ELISA application (Ngan, J.Y.G., et al. (2021). Proc Natl Acad Sci USA. 118(12):e2006717118).
ELISA Analysis: Various dilutions of this antibody detected recombinant GTPase HfIX protein.
Immunoprecipitation Analysis: A representative lot immunoprecipitated GTPase HfIX in Immunoprecipitation application (Ngan, J.Y.G., et al. (2021). Proc Natl Acad Sci USA. 118(12):e2006717118).
Dot Blot: A representative lot detected HfIX in Dot Blot applications (Ngan, J.Y.G., et al. (2021). Proc Natl Acad Sci USA. 118(12):e2006717118).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone 49 is a mouse monoclonal antibody that detects GTPase High frequency of lysogenization X (HflX).
Physical form
Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
Preparation Note
1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Recommended storage: +2°C to +8°C.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
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.
