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

Product name

Anti-STING antibody [EPR25090-107]
See all STING primary antibodies
Description

Rabbit monoclonal [EPR25090-107] to STING
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, IHC-Fr, WB, IPmore details
Species reactivity

Reacts with: Mouse, Rat
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: C2C12, RAW264.7, J774A.1, and A20 whole cell lysates; Mouse spleen tissue lysate; Rat spleen tissue lysate. IHC-P: Mouse spleen, liver and breast cancer tissues; Rat spleen and liver tissues. IHC-Fr: Mouse spleen tissue; Rat spleen tissue. ICC/IF: J774A.1 and RAW 264.7 cells. Flow cyt: RAW 264.7 and J774A.1 cells. IP: RAW264.7 whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR25090-107
Isotype

IgG
Research areas

Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab288157 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/500.
ICC/IF		1/50.
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr		1/500.
WB		1/1000. Detects a band of approximately 33, 35 kDa (predicted molecular weight: 42 kDa).
IP		1/30.

Notes
Flow Cyt (Intra) 1/500.
ICC/IF 1/50.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/500.
WB 1/1000. Detects a band of approximately 33, 35 kDa (predicted molecular weight: 42 kDa).
IP 1/30.

Function

Facilitator of innate immune signaling that promotes the production of type I interferon (IFN-alpha and IFN-beta). Innate immune response is triggered in response to non-CpG double-stranded DNA from viruses and bacteria delivered to the cytoplasm. Able to activate both NF-kappa-B and IRF3 transcription pathways to induce expression of type I interferon and exert a potent anti-viral state following expression. May be involved in translocon function, the translocon possibly being able to influence the induction of type I interferons. May be involved in transduction of apoptotic signals via its association with the major histocompatibility complex class II (MHC-II). Mediates death signaling via activation of the extracellular signal-regulated kinase (ERK) pathway.
Tissue specificity

Ubiquitously expressed.
Sequence similarities

Belongs to the TMEM173 family.
Post-translational
modifications

Phosphorylated on tyrosine residues upon MHC-II aggregation (By similarity). Phosphorylated on Ser-358 by TBK1, leading to activation and production of IFN-beta.
Ubiquitinated. 'Lys-63'-linked ubiquitination mediated by TRIM56 at Lys-150 promotes homodimerization and recruitment of the antiviral kinase TBK1 and subsequent production of IFN-beta. 'Lys-48'-linked polyubiquitination at Lys-150 occurring after viral infection is mediated by RNF5 and leads to proteasomal degradation.
Cellular localization

Endoplasmic reticulum membrane. Mitochondrion outer membrane. Cell membrane. Cytoplasm > perinuclear region. In response to double-stranded DNA stimulation, relocalizes to perinuclear region, where the kinase TBK1 is recruited.
Target information above from: UniProt accession Q86WV6 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 72512 Mouse
- Entrez Gene: 498840 Rat
- SwissProt: Q3TBT3 Mouse
- Unigene: 45995 Mouse
Alternative names
- endoplasmic reticulum IFN stimulator antibody
- Endoplasmic reticulum interferon stimulator antibody
- ERIS antibody
- FLJ38577 antibody
- hMITA antibody
- hSTING antibody
- Mediator of IRF3 activation antibody
- MITA antibody
- Mitochondrial mediator of IRF3 activation antibody
- MPYS antibody
- N terminal methionine proline tyrosine serine plasma membrane tetraspanner antibody
- NET23 antibody
- Stimulator of interferon genes antibody
- Stimulator of interferon genes protein antibody
- STING antibody
- TM173_HUMAN antibody
- Tmem173 antibody
- Transmembrane protein 173 antibody
see all

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
Western blot - Anti-STING antibody [EPR25090-107] (ab288157)

All lanes : 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 (ab97051) at 1/100000 dilution

Predicted band size: 42 kDa
Observed band size: 33, 35 kDa why is the actual band size different from the predicted?

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

Exposure time: 81 seconds
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.
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.
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
Western blot - Anti-STING antibody [EPR25090-107] (ab288157)

All lanes : 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 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 42 kDa
Observed band size: 33, 35 kDa why is the actual band size different from the predicted?

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

Exposure time: 26 seconds
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.
Western blot - Anti-STING antibody [EPR25090-107] (ab288157)

All lanes : Anti-STING antibody [EPR25090-107] (ab288157) at 1/1000 dilution

Lane 1 : C2C12 (mouse myoblasts myoblast) whole cell lysate
Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : J774A.1 (mouse reticum cell sarcoma monocyte macrophage) whole cell lysate
Lane 4 : A20 (mouse reticum sarcoma b lymphocyte) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

Predicted band size: 42 kDa
Observed band size: 33.35 kDa why is the actual band size different from the predicted?

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

Exposure time: 5.5 seconds

Lysates/proteins at 20 µg per lane.
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.
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.
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.
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
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
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
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
Anti-STING antibody [EPR25090-107] (ab288157)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As

Post-translational
modifications