Anti-STING antibody [EPR13130]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] (AB181125)

Overlay histogram showing THP-1 cells fixed in 4% PFA and stained with purified 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).

Alexa Fluor^® 488 (ab198950) and R-PE (ab208874) conjugated versions are available for this clone.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-STING antibody [EPR13130] (AB181125)

Intracellular Flow Cytometry analysis of THP1 cells using unpurified ab181125at a 1/10 dilution (red) or a Rabbit monoclonal IgG (negative) (green). Goat anti rabbit IgG (FITC) secondary used at a 1/150 dilution.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR13130] (AB181125)

Immunocytochemistry/immunofluorescence analysis of THP-1 (human monocytic leukemia monocyte) cells labelling TMEM173 with ab181125 at 10 μg/mL. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red). Nuclear DNA was labelled with DAPI (blue).
Confocal image showing cytoplasmic staining in THP-1 cells.

• IP

Lab

Immunoprecipitation - Anti-STING antibody [EPR13130] (AB181125)

Immunoprecipitation of STING1 in THP-1 cells. Lysates were prepared and immunoprecipitation was performed using 2 µg of 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] (ab181125) at 2 µg

All lanes:

THP-1 cells

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• WB

Lab

Western blot - Anti-STING antibody [EPR13130] (AB181125)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-STING antibody [EPR13130] (ab181125) at 1/2000 dilution

All lanes:

HEK293 cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 42 kDa

Observed band size: 37 kDa

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• WB

Lab

Western blot - Anti-STING antibody [EPR13130] (AB181125)

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 °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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - 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 (<a href='/en-us/products/cell-lines/human-tmem173-knockout-thp-1-cell-line-ab270493'>ab270493</a>)

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

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• WB

Lab

Western blot - Anti-STING antibody [EPR13130] (AB181125)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-STING antibody [EPR13130] (ab181125) at 1/10000 dilution

Lane 1:

THP-1 cell lysate at 20 µg

Lane 2:

HACAT cell lysate at 20 µg

Lane 3:

human spleen lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 42 kDa

Observed band size: 37 kDa

false

• WB

Lab

Western blot - Anti-STING antibody [EPR13130] (AB181125)

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] (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

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• WB

Supplier Data

Western blot - Anti-STING antibody [EPR13130] (AB181125)

All lanes:

Western blot - Anti-STING antibody [EPR13130] (ab181125) at 1/2000 dilution

Lane 1:

THP1 lysate at 20 µg

Lane 2:

HACAT lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 42 kDa

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