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16029
Mouse-Reactive STING Pathway Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Mouse-Reactive STING Pathway Antibody Sampler Kit #16029

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Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 1

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).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 2

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).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 3

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).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 4

Western blot analysis of HCT116 cell extracts, untreated (-) or TBK1/NAK knock-out (+), using TBK/NAK (D1B4) Rabbit mAb Antibody #3504 (upper) or β-actin (D6A8) Rabbit mAb #8457 (lower).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 5

Flow cytometric analysis of THP-1 cells differentiated with TPA (80nM, 4 days) #9905, untreated (blue) or treated with LPS (1 ng/mL, 1 hr; green) #14011 using Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 6

Western blot analysis of extracts from control HeLa cells (lane 1) or IRF-3 knockout HeLa cells (lane 2) using IRF-3 (D83B9) Rabbit mAb, #4302 (upper) or β-actin (13E5) Rabbit mAb, #4970 (lower). The absence of signal in the IRF-3 knockout HeLa cells confirms specificity of the antibody for IRF-3.

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 7

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).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 8

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.

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 9

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.

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 10

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.

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 11

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

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 12

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

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 13

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

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 14

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).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 15

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.

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 16

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).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 17

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).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 18

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

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 19

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).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 20

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).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 21

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).

Mouse-Reactive STING Pathway Antibody Sampler Kit: Image 22

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).

To Purchase # 16029T
Product # Size Price
16029T
1 Kit  (7 x 20 µl) N/A

Product Description

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.

Specificity / Sensitivity

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.

Source / Purification

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.

Background

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.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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