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16029
Mouse-Reactive STING Pathway Antibody Sampler Kit
Primary Antibodies

Mouse-Reactive STING Pathway Antibody Sampler Kit, UniProt ID Q14653, Entrez ID 214763 #16029

Western Blotting Image 1 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of extracts from various cell lines using STING (D1V5L) Rabbit mAb (Rodent Preferred).

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Western Blotting Image 2 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of extracts from Raw 264.7 cells, untransfected (-) or transfected with poly(dA:dT) (5 μg/mL, 3 hr; +), using Phospho-STING (Ser365) (D8F4W) Rabbit mAb (upper), STING (D1V5L) Rabbit mAb (Rodent Preferred) #50494 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).

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Western Blotting Image 3 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of extracts from 3T3, Neuro-2a, and C2C12 cells using cGAS (D3O8O) Rabbit mAb (Mouse Specific).

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Western Blotting Image 4 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of extracts from various cell lines using TBK1/NAK (D1B4) Rabbit mAb.

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Western Blotting Image 5 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of extracts from THP-1 cells differentiated with TPA #4174 (80 nM, overnight) followed by treatment with LPS (1 μg/ml), up to 24h, using Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb (upper), or total TBK1/NAK (D1B4) Rabbit mAb #3504 (lower).

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Western Blotting Image 6 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of HeLa cell extracts, untreated (-) or IRF-3 knock-out (+), using IRF-3 (D83B9) Rabbit mAb, #4302 (upper) or β-actin (13E5) Rabbit mAb #4970 (lower).

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Western Blotting Image 7 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of HT-29 cells, untransfected (-) or transfected with Poly (I:C) (2.5 μg/ml, 6 hr; +), using Phospho-IRF-3 (Ser396) (D6O1M) Rabbit mAb (upper) or IRF-3 (D6I4C) XP® Rabbit mAb #11904 (lower).

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Western Blotting Image 8 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of adipocytes from wild type (WT) mice, untransfected (-) or transfected with Poly (I:C) (2.5 μg/ml, 6 hr; +), and adipocytes from IRF-3 (-/-) mice, untransfected (-) or transfected with Poly (I:C) (2.5 μg/ml, 6 hr; +), using Phospho-IRF-3 (Ser396) (D6O1M) Rabbit mAb (upper), IRF-3 (D83B9) Rabbit mAb #4302 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).

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Flow Cytometry Image 9 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Flow cytometric analysis of adipocytes from wild type mice, untransfected (A) or transfected with Poly (I:C) (2.5 µg/ml, 6 hr; B), and adipocytes from IRF-3 (-/-) mice, untransfected (C) or transfected with Poly (I:C) (2.5 µg/ml, 6 hr; D), using Phospho-IRF-3 (Ser396) (D6O1M) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

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Western Blotting Image 10 - Mouse-Reactive STING Pathway Antibody Sampler Kit

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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IP Image 11 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Immunoprecipitation of STING from Raw 264.7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is STING (D1V5L) Rabbit mAb (Rodent Preferred). Western blot analysis was performed using STING (D1V5L) Rabbit mAb (Rodent Preferred).

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Western Blotting Image 12 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western Blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length mouse cGAS protein (mcGAS-Myc/DDK; +), using cGAS (D3O8O) Rabbit mAb (Mouse Specific).

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Western Blotting Image 13 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of extracts from THP-1 cells differentiated with TPA #4174 (80 nM, overnight) followed by treatment with LPS (1 μg/ml, 1 hour), with or without phosphatase treatment using Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb (upper), or total TBK1/NAK (D1B4) Rabbit mAb #3504 (lower).

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Western Blotting Image 14 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of extracts from COS-7 cells, mock transfected or transfected with human, mouse or rat IRF-3 constructs, using IRF-3 (D83B9) Rabbit mAb. The human and mouse constructs contain epitope tags.

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IP Image 15 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Immunoprecipitation of cGAS from C2C12 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is cGAS (D3O8O) Rabbit mAb (Mouse Specific). Western blot analysis was performed using cGAS (D3O8O) Rabbit mAb (Mouse Specific).

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Flow Cytometry Image 16 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Flow cytometric analysis of THP-1 cells differentiated with TPA #9905, untreated (blue) or LPS-treated (green), using Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb.

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Western Blotting Image 17 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of extracts from various cell lines using IRF-3 (D83B9) Rabbit mAb.

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Flow Cytometry Image 18 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Flow cytometric analysis of HT-29 cells, untransfected (blue) or transfected with Poly (I:C) (2.5 μg/ml, 6 hr; green), using Phospho-IRF-3 (Ser396) (D6O1M) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

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IF-IC Image 19 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Confocal immunofluorescent analysis of THP-1 cells differentiated with TPA #4174 (80nM, overnight) (left), followed by treatment with LPS (1μg/ml, 1 hour) (center) or LPS with λ phosphatase treatment (right) using Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 Phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Western Blotting Image 20 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Western blot analysis of extracts from differentiated THP-1 cells, untreated or treated with LPS (1 µg/ml, 1 hour), using Phospho-IRF-3 (Ser396) (4D4G) Rabbit mAb #4947 (upper) or IRF-3 (D83B9) Rabbit mAb (lower).

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IF-IC Image 21 - Mouse-Reactive STING Pathway Antibody Sampler Kit

Confocal immunofluorescent analysis of HT-29 cells, untransfected (left) or transfected with Poly (I:C) (2.5 μg/ml, 6 hr; right), using Phospho-IRF-3 (Ser396) (D6O1M) Rabbit mAb (green) and EpCAM (VU1D9) Mouse mAb (Alexa Fluor® 555 Conjugate) #5488 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
STING (D1V5L) Rabbit mAb (Rodent Preferred) 50494 20 µl
  • WB
  • IP
H M R 33, 35 Rabbit IgG
Phospho-STING (Ser365) (D8F4W) Rabbit mAb 72971 20 µl
  • WB
M 41 Rabbit IgG
cGAS (D3O8O) Rabbit mAb (Mouse Specific) 31659 20 µl
  • WB
  • IP
M 62 Rabbit IgG
TBK1/NAK (D1B4) Rabbit mAb 3504 20 µl
  • WB
  • IP
H M R Mk 84 Rabbit 
Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb 5483 20 µl
  • WB
  • IP
  • IF
  • F
H M 84 Rabbit IgG
IRF-3 (D83B9) Rabbit mAb 4302 20 µl
  • WB
  • IP
H M R Mk 45-55 Rabbit IgG
Phospho-IRF-3 (Ser396) (D6O1M) Rabbit mAb 29047 20 µl
  • WB
  • IP
  • IF
  • F
H M R 45-55 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Mouse-Reactive STING Pathway Antibody Sampler Kit provides an economical means of detecting activation and expression of key components of the STING pathway using phospho-specific and control antibodies. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Each antibody in the Mouse-Reactive STING Pathway Antibody Sampler Kit detects endogenous levels of its target protein. Phospho-STING (Ser365) (D8W4F) Rabbit mAb recognizes endogenous levels of STING protein only when phosphorylated at Ser365. Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb detects endogenous levels of TBK1 only when phosphorylated at Ser172. Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb may cross-react with phospho-IKKε. Phospho-IRF-3 (Ser396) (D6O1M) Rabbit mAb recognizes endogenous levels of IRF-3 protein only when phosphorylated at Ser396.

Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to residues surrounding Ala66 of mouse cGAS protein, Ser645 of human TBK1/NAK, residues near the carboxyl terminus of human IRF-3, and recombinant mouse STING protein. Phosphorylation-specific monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding residues surrounding Ser365 of mouse STING protein, Ser172 of human TBK1, and Ser396 of human IRF-3.

Stimulator of interferon genes (STING, TMEM173, MITA) is a transmembrane adaptor protein that is a critical component of the cellular innate immune response to pathogenic cytoplasmic DNA (1,2). STING is a ubiquitously expressed protein found predominantly in the ER (1). The enzyme cGAMP synthase (cGAS) produces the second messenger cyclic-GMP-AMP (cGAMP) in response to cytoplasmic DNA (3,4). cGAMP binds and activates STING (3,4). In addition, detection of cytoplasmic DNA by nucleic acid sensors, including DDX41 or IFI16, results in STING activation (5,6). Following activation, STING translocates with TBK1 to perinuclear endosomes and gets phosphorylated by ULK1 at Ser366 (Ser365 in mouse) (7, 8). The TBK1 kinase phosphorylates and activates IRF-3 and NF-κB, which leads to the induction of type I interferon and other immune response genes (1,2,7).

  1. Ishikawa, H. and Barber, G.N. (2008) Nature 455, 674-8.
  2. Zhong, B. et al. (2008) Immunity 29, 538-50.
  3. Sun, L. et al. (2013) Science 339, 786-91.
  4. Wu, J. et al. (2013) Science 339, 826-30.
  5. Zhang, Z. et al. (2011) Nat Immunol 12, 959-65.
  6. Unterholzner, L. et al. (2010) Nat Immunol 11, 997-1004.
  7. Ishikawa, H. et al. (2009) Nature 461, 788-92.
  8. Konno, H. et al. (2013) Cell 155, 688-98.
Entrez-Gene Id
214763 , 3661 , 72512 , 29110
Swiss-Prot Acc.
Q8C6L5 , Q14653 , Q3TBT3 , Q9UHD2
For Research Use Only. Not For Use In Diagnostic Procedures.

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