Knockout Validated, Multiplex Immunofluorescence, Immunohistochemistry, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, COMET

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2922D expressed in HEK293

Format

BSA Free

Concentration

1.0 mg/ml

View all STING/TMEM173 Antibodies »

Product Specifications

Immunogen

Partial recombinant protein made to amino acids 215-379 of human TMEM173/STING (UniProt Q86WV6).

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for STING/TMEM173 Antibody (2922D) - BSA Free

Detection of STING/TMEM173 in Human Brain Cortex (Cerebrum) via seqIF™ staining on COMET™

STING/TMEM173 was detected in immersion fixed paraffin-embedded sections of human brain Cortex (Cerebrum) using Rabbit Anti-Human STING/TMEM173, Monoclonal Antibody (Catalog #NBP3-18816) at 10ug/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.The image is attached.

Detection of STING in Frozen Mouse Hippocampus via seqIF™ staining on COMET™

STING was detected in frozen sections of mouse hippocampus using Rabbit Anti-Mouse STING Monoclonal Antibody (Catalog # NBP3-18816) at 10ug/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent preprocessing by incubating tissue with Multi Staining Buffer (Lunaphore Catalog # BU06) for 5minutes at room temperature followed by a 20-minute incubation in Tris-Buffered Saline + 0.2% Triton at room temperature. Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.

Detection of STING in Frozen Mouse Cerebral Cortex via seqIF™ staining on COMET™

STING was detected in frozen sections of mouse cerebral cortex using Rabbit Anti-Mouse STING Monoclonal Antibody (Catalog # NBP3-18816) at 10ug/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent preprocessing by incubating tissue with Multi Staining Buffer (Lunaphore Catalog # BU06) for 5minutes at room temperature followed by a 20-minute incubation in Tris-Buffered Saline + 0.2% Triton at room temperature. Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.

Detection of STING in Frozen Mouse Choroid Plexus via seqIF™ staining on COMET™

STING was detected in frozen sections of mouse choroid plexus using Rabbit Anti-Mouse STING Monoclonal Antibody (Catalog # NBP3-18816) at 10ug/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent preprocessing by incubating tissue with Multi Staining Buffer (Lunaphore Catalog # BU06) for 5minutes at room temperature followed by a 20-minute incubation in Tris-Buffered Saline + 0.2% Triton at room temperature. Tissue was stained using the Alexa Fluor™ Plus 647 Goat anti-Rabbit IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RB) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm. Protocol available in COMET™ Panel Builder.

Western Blot: STING/TMEM173 Antibody (2922D) [NBP3-18816]

Western Blot: STING/TMEM173 Antibody (2922D) [NBP3-18816] - Western blot shows cell lysates K562, THP-1, C2C12, and C6. Membrane was probed with 1 ug/mL of (Catalog # NBP3-18816) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for STING/TMEM173 at approximately 42 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1. Internally validated Western blot.

Immunocytochemistry/ Immunofluorescence: STING/TMEM173 Antibody (2922D) [NBP3-18816]

Immunocytochemistry/Immunofluorescence: STING/TMEM173 Antibody (2292D) [NBP3-18816] - STING/TMEM173 was detected in immersion fixed U937 human myeloid leukaemia cell line but is not detected in Daudi cell line using Rabbit Anti-Human STING/TMEM173 Monoclonal Antibody (Catalog # NBP3-18816) at 1 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights(TM) 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue).

Immunohistochemistry: STING/TMEM173 Antibody (2922D) [NBP3-18816]

Immunohistochemistry: STING/TMEM173 Antibody (2292D) [NBP3-18816] - STING/TMEM173 was detected in immersion fixed paraffin-embedded sections of mouse lung tissue using Rabbit Anti-Human STING/TMEM173 Monoclonal Antibody at 0.5 ug/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody. Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to pneumocytes and alveolar cells. Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western: STING/TMEM173 Antibody (2922D) [NBP3-18816]

Simple Western: STING/TMEM173 Antibody (2922D) [NBP3-18816] - Simple Western Lane view shows K562 and THP-1 whole cell lysate (WCL). A specific band was detected for hSTING/TMEM173 antibody (NBP3-18816) at approximately 44 kDa (as indicated) using 10 ug/mL of hSTING/TMEM173 antibody. This experiment was conducted under reducing conditions.

Western Blot Shows Human STING/TMEM173 Specificity Using Knockout Cell Line.

Western blot shows lysates of THP-1 cell line and STING/TMEM173 knockout THP-1 cell line (KO). Nitrocellulose membrane was probed with STING/TMEM173 Antibody (Catalog # NBP3-18816) followed by HRP-conjugated secondary antibody. A specific band was detected for STING/TMEM173 at approximately 41 kDa (as indicated) in the parental THP-1 cell line, but is not detectable in knockout THP-1 cell line. Primary antibody dilution used: 1/1000. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Detection of STING/TMEM173 by Immunoprecipitation.

PMA-treated THP-1 lysates were prepared and immunoprecipitation was performed using 2.0 µg of STING/TMEM173 Antibody (Catalog # NBP3-18816) pre-coupled to Dynabeads protein A. Immunoprecipitated STING/TMEM173 was detected with NBP3-18816. For western blot, NBP3-18816 was used at 1/1000. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

STING/TMEM173 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

PMA-treated THP-1 WT and STING/TMEM173 KO cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with STING/TMEM173 Antibody (Catalog # NBP3-18816) and with an Alexa-fluor 555 coupled secondary antibody including DAPI. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative images of the blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Primary antibody dilution used: 1/1000. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

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Applications for STING/TMEM173 Antibody (2922D) - BSA Free

Application

Recommended Usage

Immunocytochemistry/ Immunofluorescence

3 ug/ml

Immunohistochemistry

0.5 ug/ml

Immunoprecipitation

Validated for Immunoprecipitation from YCharOS Inc. (ycharos.com)

Knockout Validated

Validated for Knockout from YCharOS Inc. (ycharos.com)

Multiplex Immunofluorescence

10 ug/mL

Western Blot

1 - 2 ug/ml

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.

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 Antibody (2922D) - BSA Free

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for STING/TMEM173 Antibody (2922D) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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STING/TMEM173 Antibody (2922D) - BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-18816

Key Product Details

Validated by

Species Reactivity

Applications

Label

Antibody Source

Format

Concentration

View all STING/TMEM173 Antibodies »

Product Specifications

Immunogen

Clonality

Host

Isotype

Scientific Data Images for STING/TMEM173 Antibody (2922D) - BSA Free

Detection of STING/TMEM173 in Human Brain Cortex (Cerebrum) via seqIF™ staining on COMET™

Detection of STING in Frozen Mouse Hippocampus via seqIF™ staining on COMET™

Detection of STING in Frozen Mouse Cerebral Cortex via seqIF™ staining on COMET™

Detection of STING in Frozen Mouse Choroid Plexus via seqIF™ staining on COMET™

Western Blot: STING/TMEM173 Antibody (2922D) [NBP3-18816]

Immunocytochemistry/ Immunofluorescence: STING/TMEM173 Antibody (2922D) [NBP3-18816]

Immunohistochemistry: STING/TMEM173 Antibody (2922D) [NBP3-18816]

Simple Western: STING/TMEM173 Antibody (2922D) [NBP3-18816]

Western Blot Shows Human STING/TMEM173 Specificity Using Knockout Cell Line.

Detection of STING/TMEM173 by Immunoprecipitation.

STING/TMEM173 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

Applications for STING/TMEM173 Antibody (2922D) - BSA Free

Immunocytochemistry/ Immunofluorescence

Immunohistochemistry

Immunoprecipitation

Knockout Validated

Multiplex Immunofluorescence

Western Blot

Formulation, Preparation, and Storage

Purification

Formulation

Format

Preservative

Concentration

Shipping

Stability & Storage

Background: STING/TMEM173

Long Name

Alternate Names

Gene Symbol

Additional STING/TMEM173 Products

Product Documents for STING/TMEM173 Antibody (2922D) - BSA Free

Product Datasheets

COA

Certificate of Analysis

SDS

Product Specific Notices for STING/TMEM173 Antibody (2922D) - BSA Free