STING/TMEM173 Overexpression Lysate

Name: STING/TMEM173 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-17038

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-17038

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for STING/TMEM173 Overexpression Lysate

STING/TMEM173 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC208418

Accession#: NM_198282

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

42 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for STING/TMEM173 Overexpression Lysate

Western Blot: STING/TMEM173 Overexpression Lysate [NBL1-17038]

Western Blot: STING/TMEM173 Overexpression Lysate [NBL1-17038] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right: STING/TMEM173 Overexpression Lysate

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: STING/TMEM173

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

Long Name

Stimulator of Interferon Genes Protein/Transmembrane protein 173

Alternate Names

ERIS, MITA, MPYS, NET23, TMEM173

Gene Symbol

STING1

Additional STING/TMEM173 Products

Product Documents for STING/TMEM173 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for STING/TMEM173 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

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.

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