Anti-STING antibody [EPR25090-107]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR25090-107] (AB288157)

Immunohistochemical analysis of paraffin-embedded (A) Liver tissue from wild-type C57BL/6JGpt mice (B) Liver tissue from STING knockout mice labeling STING with ab288157 at 1/5000 followed by a ready to use LeicaDS9800 (Bond®, Polymer Refine Detection). Counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.Positive staining on (A) Liver tissue from wild-type C57BL/6JGpt mice and no staining on (B) Liver tissue from STING knockout mice. The section was incubated with ab288157 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Sting1-KO homozygous mice (Strain ID : T012747).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR25090-107] (AB288157)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling STING with ab288157 at 1/2000 (0.279 ug/ml) followed by a ready to use LeicaDS9800 (Bond^®, Polymer Refine Detection). Positive staining on kupffer cells in rat liver (PMID 30561388). The section was incubated with ab288157 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond®, Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR25090-107] (AB288157)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 cells labelling STING with ab288157 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing positive cytoplasmic staining in RAW 264.7 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR25090-107] (AB288157)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling STING with ab288157 at 1/2000 (0.279 ug/ml) followed by a ready to use LeicaDS9800 (Bond®, Polymer Refine Detection). Positive staining on mouse spleen. The section was incubated with ab288157 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^®, Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-STING antibody [EPR25090-107] (AB288157)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat spleen (fresh) tissue labeling STING with ab288157 at 1/500 (1.114 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Positive staining on rat spleen is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR25090-107] (AB288157)

Immunohistochemical analysis of paraffin-embedded (A) Liver tissue from wild-type C57BL/6JGpt mice (B) Liver tissue from STING knockout mice labeling STING with ab288157 at 1/5000 followed by a ready to use LeicaDS9800 (Bond®, Polymer Refine Detection). Counterstained with Hematoxylin. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Positive staining on (A) Liver tissue from wild-type C57BL/6JGpt mice and no staining on (B) Liver tissue from STING knockout mice. The section was incubated with ab288157 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Sting1-KO homozygous mice (Strain ID : T012747).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-STING antibody [EPR25090-107] (AB288157)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) cells labelling STING with ab288157 at 1/500 dilution (0.1ug)(Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-STING antibody [EPR25090-107] (AB288157)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 cells labelling STING with ab288157 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing positive cytoplasmic staining in J774A.1 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR25090-107] (AB288157)

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labeling STING with ab288157 at 1/2000 (0.279 ug/ml) followed by a ready to use LeicaDS9800 (Bond^®, Polymer Refine Detection). Positive staining on mouse breast cancer. The section was incubated with ab288157 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^®, Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-STING antibody [EPR25090-107] (AB288157)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen (fresh) tissue labeling STING with ab288157 at 1/500 (1.114 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (Green). Positive staining on mouse spleen is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-STING antibody [EPR25090-107] (AB288157)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling STING with ab288157 at 1/500 dilution (0.1ug)(Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR25090-107] (AB288157)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling STING with ab288157 at 1/2000 (0.279 ug/ml) followed by a ready to use LeicaDS9800 (Bond^®, Polymer Refine Detection). Positive staining on kupffer cells in mouse liver (PMID 30561388). The section was incubated with ab288157 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^®, Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STING antibody [EPR25090-107] (AB288157)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling STING with ab288157 at 1/2000 (0.279 ug/ml) followed by a ready to use LeicaDS9800 (Bond®, Polymer Refine Detection) was used. Positive staining on rat spleen. The section was incubated with ab288157 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond®, Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Lab

Immunoprecipitation - Anti-STING antibody [EPR25090-107] (AB288157)

STING was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab288157 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288157 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 ug

Lane 2 : ab288157 IP in RAW264.7 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288157 in RAW264.7 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 7.75 seconds

All lanes:

Immunoprecipitation - Anti-STING antibody [EPR25090-107] (ab288157)

Predicted band size: 42 kDa

Observed band size: 33 kDa,33.35 kDa,35 kDa

false

• WB

Lab

Western blot - Anti-STING antibody [EPR25090-107] (AB288157)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 81 seconds

All lanes:

Western blot - Anti-STING antibody [EPR25090-107] (ab288157) at 1/1000 dilution

Lane 1:

Rat spleen tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 33 kDa,35 kDa

false

• WB

Lab

Western blot - Anti-STING antibody [EPR25090-107] (AB288157)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 26 seconds

All lanes:

Western blot - Anti-STING antibody [EPR25090-107] (ab288157) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 33 kDa,33.35 kDa,35 kDa

false

• WB

Lab

Western blot - Anti-STING antibody [EPR25090-107] (AB288157)

Loading control : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1/200000) (36KDa) Blocking buffer and concentration : 5% NFDM/TBST The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Sting1-KO homozygous mice (Strain ID : T012747).

Lanes 1 - 6:

Western blot - Anti-STING antibody [EPR25090-107] (ab288157) at 1/1000 dilution

Lanes 1 - 6:

Western blot - Anti-STING antibody [EPR25090-107] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/sting-antibody-epr25090-107-bsa-and-azide-free-ab288164'>ab288164</a>)

Lane 1:

Wild-type mouse spleen tissue lysate (male)

Lanes 2 - 3:

STING knockout spleen tissue lysate (male)

Lane 4:

Wild-type mouse liver tissue lysate (male)

Lanes 5 - 6:

STING knockout liver tissue lysate (male)

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-STING antibody [EPR25090-107] (AB288157)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 5.5 seconds

Lysates/proteins at 20 μg per lane.

All lanes:

Western blot - Anti-STING antibody [EPR25090-107] (ab288157) at 1/1000 dilution

Lane 1:

C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

J774A.1 (mouse reticum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg

Lane 4:

A20 (mouse reticum sarcoma b lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 42 kDa

Observed band size: 33.35 kDa

false