STING Recombinant Rabbit Monoclonal Antibody [JM03-47]
cat.: ET1705-68
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Rat
Applications: WB, IHC-P, IF-Cell, FC, IF-Tissue, IP
Clonality: Monoclonal
Clone number: JM03-47
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 42 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human TMEM173 aa 220-379 / 379.
Positive control: THP-1 cell lysate, SW620 cell lysate, human lung tissue lysate, THP-1, human tonsil tissue, rat lung tissue.
Subcellular location: Endoplasmic reticulum membrane, Cytoplasm, perinuclear region, Endoplasmic reticulum-Golgi intermediate compartment membrane, Golgi apparatus membrane, Cytoplasmic vesicle, autophagosome membrane, Mitochondrion outer membrane, Cell membrane.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC
  IF-Tissue
  IP

1:500-1:1,000
1:1,000
1:100
1:1,000
1:200
1-2μg/sample
Uniprot #: SwissProt: Q86WV6 Human | F1M391 Rat
Alternative names: endoplasmic reticulum IFN stimulator Endoplasmic reticulum interferon stimulator ERIS FLJ38577 hMITA hSTING Mediator of IRF3 activation MITA Mitochondrial mediator of IRF3 activation MPYS N terminal methionine proline tyrosine serine plasma membrane tetraspanner NET23 Stimulator of interferon genes Stimulator of interferon genes protein STING TM173_HUMAN Tmem173 Transmembrane protein 173
Images
ET1705-68_1.jpg Fig1: Western blot analysis of STING on different lysates with Rabbit anti-STING antibody (ET1705-68) at 1/1,000 dilution.

Lane 1: THP-1 cell lysate (15 µg/Lane)
Lane 2: SW620 cell lysate (15 µg/Lane)
Lane 3: Human lung tissue lysate (30 µg/Lane)

Predicted band size: 42 kDa
Observed band size: 35 kDa

Exposure time: 1 minute 2 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-68) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1705-68_2.jpg Fig2: Immunocytochemistry analysis of THP-1 cells labeling STING with Rabbit anti-STING antibody (ET1705-68) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-STING antibody (ET1705-68) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1705-68_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-STING antibody (ET1705-68) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-68) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1705-68_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-STING antibody (ET1705-68) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-68) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1705-68_5.jpg Fig5: Flow cytometric analysis of THP-1 cells labeling STING.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1705-68, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1705-68_6.jpg Fig6: STING was immunoprecipitated from 0.2 mg THP-1 cell lysate with ET1705-68 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1705-68 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: THP-1 cell lysate (input)
Lane 2: ET1705-68 IP in THP-1 cell lysate
Lane 3: Rabbit IgG instead of ET1705-68 in THP-1 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.