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38866
Human-Reactive STING Pathway Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit
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Human-Reactive STING Pathway Antibody Sampler Kit #38866

Citations (6)
Simple Western™ analysis of lysates (1 mg/mL) from HeLa cells using IRF-3 (D6I4C) XP ® Rabbit mAb #11904. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from control HeLa cells (lane 1) or IRF-3 knockout HeLa cells (lane 2) using IRF-3 (D6I4C) XP® Rabbit mAb, #11904 (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.
Western blot analysis of extracts from various cell lines using STING (D2P2F) Rabbit mAb.
Western blot analysis of extracts from various cell lines using cGAS (D1D3G) Rabbit mAb.
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).
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).
Western blot analysis of extracts from THP-1 cells differentiated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (80 nM, 16 hr) and then untransfected (-) or transfected with poly(dA:dT) (5 μg/mL, 3 hr; +) using Phospho-STING (Ser366) (E9A9K) Rabbit mAb (upper), STING (D2P2F) Rabbit mAb #13647 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
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).
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.
Western blot analysis of extracts from various cell lines using IRF-3 (D6I4C) XP® Rabbit mAb.
Western blot analysis of extracts from 293T cells, mock transfected (-), transfected with a construct expressing human STING protein (hSTING; +), or transfected with a construct expressing mouse STING protein (mSTING; +), using STING (D2P2F) Rabbit mAb.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc-tagged full-length human cGAS protein (hcGAS-Myc; +), using cGAS (D1D3G) Rabbit mAb.
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).
Western blot analysis of extracts from various cell lines using TBK1/NAK (D1B4) Rabbit mAb.
Immunoprecipitation of Phospho-STING (Ser366) protein from THP-1 extracts differentiated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (80 nM, 16 hr) and then transfected with poly(dA:dT) (5 ug/mL, 3 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-STING (Ser366) (E9A9K) Rabbit mAb. Western blot analysis was performed using Phospho-STING (Ser366) (E9A9K) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678, followed by Anti-mouse IgG, HRP-linked Antibody #7076 were used as the secondary antibodies.
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).
Immunoprecipitation of IRF-3 from THP-1 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or IRF-3 (D6I4C) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using IRF-3 (D6I4C) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 and Anti-mouse IgG, HRP-linked Antibody #7076 were used as secondary antibodies.
Immunoprecipitation of STING from HL-60 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or STING (D2P2F) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using STING (D2P2F) Rabbit mAb.
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).
Confocal immunofluorescent analysis of THP-1 cells differentiated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (80 nM, 16 hr) and then mock-transfected (left) or transfected with poly(dA:dT) (5 μg/mL, 3 hr; right) using Phospho-STING (Ser366) (E9A9K) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
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).
Confocal immunofluorescent analysis of HT-29 cells, untransfected (left) or transfected with Poly(I:C) (2.5 μg/ml, 6 hr; right), using IRF-3 (D6I4C) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using STING (D2P2F) Rabbit mAb performed on the Leica® BOND™ Rx. 
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.
Flow cytometric analysis of THP-1 cells differentiated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (80 nM, 24 hrs), and then untransfected (blue) or transfected with poly(dA:dT) (5 ug/mL, 3 hr; green), using Phospho-STING (Ser366) (E9A9K) Rabbit mAb (solid lines) or a 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.
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.
Immunohistochemical analysis of paraffin-embedded human neuroendocrine lung carcinoma using STING (D2P2F) Rabbit mAb performed on the Leica® BOND™ Rx. 
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.
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin’s lymphoma using STING (D2P2F) Rabbit mAb performed on the Leica® BOND™ Rx. 
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using STING (D2P2F) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using STING (D2P2F) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using STING (D2P2F) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human tonsil using STING (D2P2F) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse lung using STING (D2P2F) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded Syrian hamster uterus using STING (D2P2F) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded HT-29 (left, positive) and A549 (right, negative) cell pellets using STING (D2P2F) Rabbit mAb.
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To Purchase # 38866
Cat. # Size Qty. Price
38866T		 1 Kit  (7 x 20 microliters)

Bulk & Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
STING (D2P2F) Rabbit mAb 13647 20 µl
  • WB
  • IP
  • IHC
 H M Hm 33, 35 Rabbit IgG
Phospho-STING (Ser366) (E9A9K) Rabbit mAb 50907 20 µl
  • WB
  • IP
  • IF
  • F
 H 40 Rabbit IgG
cGAS (D1D3G) Rabbit mAb 15102 20 µl
  • WB
 H 62 Rabbit IgG
TBK1/NAK (D1B4) Rabbit mAb 3504 20 µl
  • WB
  • IP
 H M R Mk 84 Rabbit IgG
Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb 5483 20 µl
  • WB
  • IP
  • IF
  • F
 H M 84 Rabbit IgG
IRF-3 (D6I4C) XP® Rabbit mAb 11904 20 µl
  • WB
  • IP
  • IF
 H Mk 50-55 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 

Product Description

The Human-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 antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Human-Reactive STING Pathway Antibody Sampler Kit detects endogenous levels of its target protein. Phospho-STING (Ser366) (E9A9K) Rabbit mAb recognizes endogenous levels of STING protein only when phosphorylated at Ser366. 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 Ala19 of human cGAS protein, Ser645 of human TBK1/NAK, Pro226 of human STING, and recombinant human IRF-3 protein. Phosphorylation-specific monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to residues surrounding Ser366 of human 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).

Pathways

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For Research Use Only. Not for Use in Diagnostic Procedures.
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