Anti-STING antibody [EPR13130] - BSA and Azide free

9 product images

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  Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR13130] - BSA and Azide free (ab227128)

  Clone EPR13130 (ab227128) has been successfully conjugated by Abcam. This image was generated using Anti-TMEM173 antibody [EPR13130] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-STING antibody [EPR13130] ab198950 for protocol details.

  Alexa Fluor® 488 Anti-STING antibody [EPR13130] ab198950 staining TMEM173 in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Alexa Fluor® 488 Anti-STING antibody [EPR13130] ab198950 at 1/200 dilution (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Western blot - Anti-STING antibody [EPR13130] - BSA and Azide free (ab227128)

  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]) 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 Human TMEM173 knockout THP-1 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 °C. 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.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-STING antibody [EPR13130] ab181125).

  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 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

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  Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] - BSA and Azide free (ab227128)

  Clone EPR13130 (ab227128) has been successfully conjugated by Abcam. This image was generated using Anti-TMEM173 antibody [EPR13130] (PE). Please refer to PE Anti-STING antibody [EPR13130] ab208874 for protocol details.
  

  Overlay histogram showing HeLa cells stained with PE Anti-STING antibody [EPR13130] ab208874 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (PE Anti-STING antibody [EPR13130] ab208874, 1/500 dilution) for 30 min at 22°C.Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycoerythrin (PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

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  Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR13130] - BSA and Azide free (ab227128)

  Clone EPR13130 (ab227128) has been successfully conjugated by Abcam. This image was generated using Anti-TMEM173 antibody [EPR13130] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-STING antibody [EPR13130] ab198952 for protocol details.

  Alexa Fluor® 647 Anti-STING antibody [EPR13130] ab198952 staining TMEM173 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with Alexa Fluor® 647 Anti-STING antibody [EPR13130] ab198952 at 1/100 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  This product also gave a positive signal under the same testing conditions in HeLacells fixed with 4% formaldehyde (10 min).

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  Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] - BSA and Azide free (ab227128)

  Overlay histogram showing THP-1 cells fixed in 4% PFA and stained with purified Anti-STING antibody [EPR13130] ab181125 at a dilution of 1 in 20 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-STING antibody [EPR13130] ab181125).
  

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  Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] - BSA and Azide free (ab227128)

  Intracellular Flow Cytometry analysis of THP1 cells using unpurified Anti-STING antibody [EPR13130] ab181125 at a 1/10 dilution (red) or a Rabbit monoclonal IgG (negative) (green). Goat anti rabbit IgG (FITC) secondary used at a 1/150 dilution.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-STING antibody [EPR13130] ab181125).

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  Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR13130] - BSA and Azide free (ab227128)

  Immunofluorescence staining of HeLa cells with purified Anti-STING antibody [EPR13130] ab181125 at a working dilution of 1/1000, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified Anti-STING antibody [EPR13130] ab181125 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-STING antibody [EPR13130] ab181125).

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  Immunoprecipitation - Anti-STING antibody [EPR13130] - BSA and Azide free (ab227128)

  This data was developed using Anti-STING antibody [EPR13130] ab181125, the same antibody clone in a different buffer formulation.

  Immunoprecipitation of STING1 in THP-1 cells. Lysates were prepared and immunoprecipitation was performed using 2 µg of Anti-STING antibody [EPR13130] ab181125 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.
  
  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: Immunoprecipitation - Anti-STING antibody [EPR13130] (Anti-STING antibody [EPR13130] ab181125) at 2 µg

  All lanes: THP-1 cells

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  Western blot - Anti-STING antibody [EPR13130] - BSA and Azide free (ab227128)

  This data was developed using Anti-STING antibody [EPR13130] ab181125, the same antibody clone in a different buffer formulation.

  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 Human TMEM173 knockout THP-1 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