STING/TMEM173 Knockout HeLa Cell Lysate

Product Discontinued

STING/TMEM173 Knockout HeLa Cell Lysate Summary

• Preparation Method: Knockout achieved by using CRISPR/Cas9,-1 bp deletion in exon3 and 2 bp insertion in exon3 and Insertion of the selection cassette in exon3
• Gene: STING1

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: You will receive 1 vial (100ug) of knockout cell lysate and 1 vial (100ug) of Parental cell lysate. Lysate can be diluted with 1X SDS sample buffer and will be stable at -20 degrees C for 12 months. Minimize freeze-thaw cycles.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.1 mg cell homogenate lyophilized in RIPA buffer made with double-knockout cell lines.
• Concentration: LYOPH
• Reconstitution Instructions: To use as WB negative control, spin down briefly and resuspend in 100 uL 1xSDS sample buffer (2% SDS, 60 mM Tris-HCl pH 6.8, 10% Glycerol, 0.02% Bromophenol blue, 60 mM beta-mercaptoethanol). Boil the lysate for 3 - 5 minutes before loading it onto gel.

Lysate Details for Array

• Type: Knockout HeLa Cell

Notes

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Validation of antibody specificity is critical and verification of antibody performance against knockout samples is one way to guarantee that an antibody recognizes a specific target. Novus' KO cell lysate can be used as a negative control for western blots and to confirm the specificity of antibodies.

Alternate Names for STING/TMEM173 Knockout HeLa Cell Lysate

• endoplasmic reticulum IFN stimulator
• Endoplasmic reticulum interferon stimulator
• ERIS
• FLJ38577
• hMITA
• hSTING
• Mediator of IRF3 activation
• MITA
• mitochondrial mediator of IRF3 activation
• MPYS
• NET23
• N-terminal methionine-proline-tyrosine-serine plasma membrane tetraspanner
• SAVI
• Stimulator of interferon genes protein
• stimulator of interferon protein
• STING
• STING-beta
• TMEM173
• transmembrane protein 173

Background

STING (stimulator of interferon genes) is encoded by the TMEM173 gene and is an adaptor molecule involved in the activation of innate immune responses to PAMPS (pathogen-associated molecular patterns) and DAMPS (damage-associated molecular patterns). STING specifically recognizes cytosolic DNA products derived from pathogens (e.g., cytomegalovirus, vaccinia virus, Listeria monocytogenes) or dead cells (1, 2). In the STING pathway, dsDNA derived from pathogens or damaged cells serves as a substrate for the enzyme cGAS (cyclic GMP-AMP synthase) which produces the second messenger cyclic GMP-AMP (cGAMP) from ATP and GTP (3, 4). Under steady-state conditions STING (theoretical molecular weight 42 kDa), a protein localizes to the ER membrane. Upon activation by dsDNA derived second messenger (cGAMP), STING translocates to the Golgi apparatus as a homodimer. Once STING has trafficked to the perinuclear region, it activates TANK binding kinase 1 (TBK1), interferon regulatory factor 3 (IRF3) and NF-kB leading to the production of cytokines (e.g., type I interferon) (2, 4). Mutations in the TMEM173 gene affecting STING expression are associated with the development of the auto-inflammatory disease SAVI (STING-associated vasculopathy with onset in infancy) (2). A novel SAVI dominant mutation in the TMEM173 human gene (V155M) leads to increased localization of STING to the Golgi and perinuclear region, indicative of an activated state (1). Hallmarks of SAVI, a rare inflammatory disease, include severe vasculitis in extremities and lung inflammation (7).

References

1. Patel, S., & Jin, L. (2019). TMEM173 variants and potential importance to human biology and disease. Genes and Immunity. https://doi.org/10.1038/s41435-018-0029-9

2. Jounai, N., Kobiyama, K., Takeshita, F., & Ishii, K. J. (2013). Recognition of damage-associated molecular patterns related to nucleic acids during inflammation and vaccination. Frontiers in Cellular and Infection Microbiology. https://doi.org/10.3389/fcimb.2012.00168

3. Xiao, T. S., & Fitzgerald, K. A. (2013). The cGAS-STING Pathway for DNA Sensing. Molecular Cell. https://doi.org/10.1016/j.molcel.2013.07.004

4. Kato, K., Omura, H., Ishitani, R., & Nureki, O. (2017). Cyclic GMP-AMP as an Endogenous Second Messenger in Innate Immune Signaling by Cytosolic DNA. Annual Review of Biochemistry. https://doi.org/10.1146/annurev-biochem-061516-044813

5. Crowl, J. T., Gray, E. E., Pestal, K., Volkman, H. E., & Stetson, D. B. (2017). Intracellular Nucleic Acid Detection in Autoimmunity. Annual Review of Immunology. https://doi.org/10.1146/annurev-immunol-051116-052331

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: STING1