Human TMEM173 knockout THP-1 cell line (ab270493)

STING1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 insertion Frameshift: 99.58%.

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Images

Western blot - Human TMEM173 knockout THP-1 cell line (AB270493), expandable thumbnail

Key facts

Cell type: THP-1
Species or organism: Human
Tissue: Blood
Form: Liquid
Knockout validation: Next Generation Sequencing, Western blot
Mutation description: Knockout achieved by CRISPR/Cas9 X = 1 insertion Frameshift: 99.58%

Alternative names

ERIS, Endoplasmic reticulum interferon stimulator, FLJ38577, MITA, MPYS, Mediator of IRF3 activation, Mitochondrial mediator of IRF3 activation, N terminal methionine proline tyrosine serine plasma membrane tetraspanner, NET23, Stimulator of interferon genes, Stimulator of interferon genes protein, TM173_HUMAN, Tmem173, Transmembrane protein 173, endoplasmic reticulum IFN stimulator, hMITA, hSTING

Key facts

Cell type: THP-1
Species or organism: Human
Tissue: Blood
Form: Liquid
Knockout validation: Next Generation Sequencing, Western blot
Mutation description: Knockout achieved by CRISPR/Cas9 X = 1 insertion Frameshift: 99.58%
Disease: Acute Monocytic Leukemia
Concentration

Properties

Gene name: STING1
Gene editing type: Knockout
Gene editing method: CRISPR technology
Knockout validation: Next Generation Sequencing, Western blot

Quality control

STR analysis: CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level: EU: 1 US: 1
Adherent/suspension: Suspension
Gender: Male

Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x10⁵ cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO₂. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x10⁵ cells/mL.Small amounts of fresh media can be added until cell number/viability improves.

Subculture guidelines

All seeding densities should be based on cell counts gained by established methods.
Cells should be seeded at 2x10⁵ - 3x10⁵ cells/mL and subcultured when they have reached 8x10⁵ cells/mL.
It is not recommended to allow the cell density to exceed 1x10⁶ cells/mL.

Culture medium

RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Storage

Shipped at conditions: Dry Ice
Appropriate short-term storage conditions: -196°C
Appropriate long-term storage conditions: -196°C

Notes

Recommended control: Human wild-type THP-1 cell line (ab271147). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

Upon thawing make banks as soon as possible and use each bank within 10-20 passages
As cell line growth can vary please attempt culture for 2 weeks from revival of initial bank before contacting the technical team.

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The stimulator of interferon genes (STING) also known as TMEM173 or MPYS is a critical transmembrane protein with a molecular weight of approximately 42 kDa. It is primarily expressed in the endoplasmic reticulum of various cell types including immune cells where it plays a central role in sensing cytosolic DNA. STING binds to cyclic dinucleotides produced by the enzyme cGAS upon recognition of aberrant DNA in the cytosol. This binding initiates activation and translocation of STING to the Golgi apparatus facilitating further signaling events.

Biological function summary

STING serves as a pivotal regulator in the innate immune response to viral and bacterial infections. It operates by forming a signaling complex with kinases and other effector proteins which subsequently leads to the activation of transcription factors such as IRF3 and NF-kB. These transcription factors then induce the expression of type I interferons and other cytokines important for mounting an effective antiviral response. The STING pathway therefore enhances the immune system's ability to detect and respond to pathogens.

Pathways

The activity of STING is integral to the cGAS-STING pathway a significant cytosolic DNA-sensing pathway involved in innate immunity. Upon activation STING interacts with TBK1 a kinase that further phosphorylates IRF3 promoting its nuclear translocation and activation. Beyond this STING also intersects with pathways involving autophagy a cellular process necessary for clearing pathogens and damaged cellular components. Through these pathways STING critically contributes to upholding cellular homeostasis and immune defense.

Associated diseases and disorders

The dysregulation of STING is linked to autoinflammatory diseases and certain cancers. Abnormal STING activation can lead to chronic inflammation a feature observed in diseases such as STING-associated vasculopathy with onset in infancy (SAVI). STING's role in cancer is also notable where its ability to activate immune cells can be harnessed in immunotherapy yet its chronic activation may promote tumorigenesis. In cancer STING often interacts with proteins like K-Ras influencing tumor growth and response to therapies.

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9 product images

Western blot - Human TMEM173 knockout THP-1 cell line (ab270493)
False colour image of Western blot: Anti-STING antibody [EPR13130] staining at 1/1000 dilution shown in green; loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution shown in red. In Western blot Anti-STING antibody [EPR13130] ab181125 was shown to bind specifically to STING. A band was observed at 37 kDa in wild-type THP-1 cell lysates with no signal observed at this size in TMEM173 knockout cell line ab270493 (knockout cell lysate Human TMEM173 knockout THP-1 cell lysate ab270516). To generate this image wild-type and TMEM173 knockout THP-1 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-STING antibody [EPR13130] (Anti-STING antibody [EPR13130] ab181125) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: TMEM173 knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human TMEM173 knockout THP-1 cell line (ab270493)
Lane 3: Human Tonsil cell lysate at 20 µg
Lane 4: Human Thymus cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 37 kDa
Western blot - Human TMEM173 knockout THP-1 cell line (ab270493)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-STING antibody [EPR13130-55] ab239074 observed at 37 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
Anti-STING antibody [EPR13130-55] ab239074 was shown to react with TMEM173 in wild-type THP-1 cells in Western blot with loss of signal observed in TMEM173 knockout cell line ab270493 (TMEM173 knockout cell lysae Human TMEM173 knockout THP-1 cell lysate ab270516). Wild-type THP-1 and TMEM173 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with Anti-STING antibody [EPR13130-55] ab239074 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-STING antibody [EPR13130-55] (Anti-STING antibody [EPR13130-55] ab239074) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: TMEM173 knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human TMEM173 knockout THP-1 cell line (ab270493)
Lane 3: Human Tonsil tissue lysate at 20 µg
Lane 4: Human Thymus tissue lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 37 kDa
Western blot - Human TMEM173 knockout THP-1 cell line (ab270493)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-STING antibody [SP339] ab227705 observed at 37 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
Anti-STING antibody [SP339] ab227705 was shown to react with TMEM173 in wild-type THP-1 cells in Western blot with loss of signal observed in TMEM173 knockout cell line ab270493 (TMEM173 knockout cell lysate Human TMEM173 knockout THP-1 cell lysate ab270516). Wild-type THP-1 and TMEM173 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with Anti-STING antibody [SP339] ab227705 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ° at a 1 in 400 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-STING antibody [SP339] (Anti-STING antibody [SP339] ab227705) at 1/400 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: TMEM173 knockout THP-1 cell lysate at 20 µg
Lane 2: Western blot - Human TMEM173 knockout THP-1 cell line (ab270493)
Lane 3: Human Tonsil tissue lysate at 20 µg
Lane 4: Human Thymus tissue lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 37 kDa
Next Generation Sequencing - Human TMEM173 knockout THP-1 cell line (ab270493)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 99.58%
Western blot - Human TMEM173 knockout THP-1 cell line (ab270493)
Anti-STING antibody [SP338] ab227704 was shown to react with STING1 in wild-type THP-1 cells in Western blot with loss of signal observed in STING1 knockout cell line ab270493. Wild-type THP-1 and STING1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-STING antibody [SP338] ab227704 overnight at 4 °C at a 1/200 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-STING antibody [SP338] (Anti-STING antibody [SP338] ab227704) at 1/200 dilution
Lane 1: Wild-type THP-1 lysate at 40 µg
Lane 2: STING1 knock-out THP-1 lysate at 40 µg
Next Generation Sequencing - Human TMEM173 knockout THP-1 cell line (ab270493)
1 bp insertion after Ser127 of the WT protein
Western blot - Human TMEM173 knockout THP-1 cell line (ab270493)
Anti-STING antibody [EPR13130] ab181125 was shown to react with STING1 in wild-type THP-1 cells in Western blot with loss of signal observed in STING1 knockout cell line ab270493. Wild-type THP-1 and STING1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-STING antibody [EPR13130] ab181125 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-STING antibody [EPR13130] (Anti-STING antibody [EPR13130] ab181125) at 1/1000 dilution
Lane 1: Wild-type THP-1 lysate at 40 µg
Lane 2: STING1 knock-out THP-1 lysate at 40 µg
Western blot - Human TMEM173 knockout THP-1 cell line (ab270493)
Anti-STING antibody [SP339] ab227705 was shown to react with STING1 in wild-type THP-1 cells in Western blot with loss of signal observed in STING1 knockout cell line ab270493. Wild-type THP-1 and STING1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-STING antibody [SP339] ab227705 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-STING antibody [SP339] (Anti-STING antibody [SP339] ab227705) at 1/1000 dilution
Lane 1: Wild-type THP-1 lysate at 40 µg
Lane 2: STING1 knock-out THP-1 lysate at 40 µg
Western blot - Human TMEM173 knockout THP-1 cell line (ab270493)
Anti-STING antibody [EPR13130-55] ab239074 was shown to react with STING1 in wild-type THP-1 cells in Western blot with loss of signal observed in STING1 knockout cell line ab270493. Wild-type THP-1 and STING1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-STING antibody [EPR13130-55] ab239074 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-STING antibody [EPR13130-55] (Anti-STING antibody [EPR13130-55] ab239074) at 1/1000 dilution
Lane 1: Wild-type THP-1 lysate at 40 µg
Lane 2: STING1 knock-out THP-1 lysate at 40 µg