Human TMEM173 knockout THP-1 cell line
- Advanced Validation
- What is this?
Be the first to review this product! Submit a review
|
(0 Publication)
- WB
Lab
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 ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution shown in red. In Western blot 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-STING antibody [EPR13130] (<a href='/en-us/products/primary-antibodies/sting-antibody-epr13130-ab181125'>ab181125</a>) 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
Predicted band size: 42 kDa
Observed band size: 37 kDa
false
- WB
Lab
Western blot - Human TMEM173 knockout THP-1 cell line (AB270493)
Lanes 1 - 4 : Merged signal (red and green). Green - ab239074 observed at 37 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
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 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 ab239074 and 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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] (<a href='/en-us/products/primary-antibodies/sting-antibody-epr13130-55-ab239074'>ab239074</a>) 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
Predicted band size: 42 kDa
Observed band size: 37 kDa
false
- WB
Lab
Western blot - Human TMEM173 knockout THP-1 cell line (AB270493)
Lanes 1 - 4 : Merged signal (red and green). Green - ab227705 observed at 37 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
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 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 ab227705 and 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-STING antibody [SP339] (<a href='/en-us/products/primary-antibodies/sting-antibody-sp339-ab227705'>ab227705</a>) 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
Predicted band size: 42 kDa
Observed band size: 37 kDa
false
- WB
Lab
Western blot - Human TMEM173 knockout THP-1 cell line (AB270493)
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 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] (<a href='/en-us/products/primary-antibodies/sting-antibody-sp339-ab227705'>ab227705</a>) 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
false
- WB
Lab
Western blot - Human TMEM173 knockout THP-1 cell line (AB270493)
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 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] (<a href='/en-us/products/primary-antibodies/sting-antibody-epr13130-55-ab239074'>ab239074</a>) 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
false
- WB
Lab
Western blot - Human TMEM173 knockout THP-1 cell line (AB270493)
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 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] (<a href='/en-us/products/primary-antibodies/sting-antibody-epr13130-ab181125'>ab181125</a>) 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
false
- WB
Lab
Western blot - Human TMEM173 knockout THP-1 cell line (AB270493)
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 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] (<a href='/en-us/products/primary-antibodies/sting-antibody-sp338-ab227704'>ab227704</a>) 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
false
- NGS
Lab
Next Generation Sequencing - Human TMEM173 knockout THP-1 cell line (AB270493)
1 bp insertion after Ser127 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human TMEM173 knockout THP-1 cell line (AB270493)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 99.58%
Complementary Products
Primary Antibodies
AB181125
Anti-STING antibody [EPR13130]
20ul selling size|KO Validated|RabMAb|Recombinant
![Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] (AB181125)](https://content.abcam.com/products/images/sting-antibody-epr13130-ab181125--flow-cytometry-intracellular-img37695.jpg)
- 4
- 1
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com