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40415
MAVS Signalosome Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit
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  3. MAVS Signalosome Antibody Sampler Kit

MAVS Signalosome Antibody Sampler Kit #40415

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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 HT-29 cells, untreated or treated with h-IFNa (10 ng/mL, 16 hr), followed by poly(I:C) (2.5 mg/mL, 7 hr) and MG-132 (#2194, 10mM, 7 hr) using Phospho-IRF-3 (Ser386) (E7J8G) XP® Rabbit mAb #37829 (upper), IRF-3 (D6I4C) XP® Rabbit mAb #11904 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Simple Western™ analysis of lysates (0.1 mg/mL) from 293T cells using TRAF2 (C192) Antibody #4724. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (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 293T cells, mock transfected (-) or transfected with a construct expressing full-length human MAVS (hMAVS; +), using MAVS (D5A9E) 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 extracts from various cell lines using TRAF3 (E8H3B) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). RPMI 8226 cells harbor a heterozygous mutation in the TRAF3 gene resulting in loss of expression. Absence of signal in RPMI 8226 cells confirms the specificity of the antibody.
Immunoprecipitation of TRAF3 protein from TF-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TRAF3 (E8H3B) Rabbit mAb. Western blot analysis was performed using TRAF3 (E8H3B) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Western blot analysis of extracts from A549 cells, untreated (-) or treated with Poly (I:C) (+), using Phospho-IRF-3 (Ser386) (E7J8G) XP® Rabbit mAb (upper) or IRF-3 (D6I4C) XP® Rabbit mAb #11904 (lower).
Western blot analysis of extracts from HCT 116 cells, either wild-type (+/+) or TBK1/NAK knockout (-/-), using TBK1/NAK (E8I3G) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from Jurkat, 293, HeLa, C2C12, and NIH/3T3 cells, using TRAF2 (C192) Antibody.
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).
Western blot analysis of extracts from varous cell lines using TRAF6 (E2K9D) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Absence of signal in NCI-H23 cells is predicted by RNAseq and confirms the specificity of the antibody.
Immunoprecipitation of TRAF6 protein from K-562 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TRAF6 (E2K9D) Rabbit mAb. Western blot analysis was performed using TRAF6 (E2K9D) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Immunoprecipitation of TRAF6 protein from A20 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TRAF6 (E2K9D) Rabbit mAb. Western blot analysis was performed using TRAF6 (E2K9D) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
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 various cell lines using MAVS (D5A9E) 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).
Confocal immunofluorescent analysis of THP-1 cells, untransfected (left) or transfected with Poly (I:C) (2.5 μg/ml, 6 hr; right), using Phospho-IRF-3 (Ser386) (E7J8G) XP® Rabbit mAb (green) and β-Actin (13E5) Rabbit mAb (Alexa Fluor® 647 Conjugate) #8584 (red). Blue = Hoechst 33342 #4082 (fluorescent DNA dye).
Western blot analysis of extracts from various cell lines using TBK1/NAK (E8I3G) Rabbit mAb (upper) 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, 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 MAVS protein from MCF7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MAVS (D5A9E) Rabbit mAb. Western blot was performed using MAVS (D5A9E) 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).
Flow cytometric analysis of THP-1 cells differentiated with TPA #4174 (80 nM, 24 hr), and then untransfected (blue) or transfected with poly(dA:dT) (5 ug/mL, 3 hr; green), using Phospho-IRF-3 (Ser 386) (E7J8G) 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.
Immunoprecipitation of TBK1/NAK from Raji cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TBK1/NAK (E8I3G) Rabbit mAb. Western blot analysis was performed using TBK1/NAK (E8I3G) Rabbit mAb.
Western blot analysis of U20S extracts from WT (left) or TBK1 KO (right) using TBK1/NAK (E8I3G) Rabbit mAb. Membranes stained with Ponceau S for total protein normalization (lower). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies, as a companion to validation data generated by CST scientists.
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).
Confocal immunofluorescent analysis of MCF7 cells showing colocalization of MAVS (D5A9E) Rabbit mAb (left, green) with the mitochondrial dye MitoTracker® Red CMXRos #9082 (middle, red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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.
Confocal immunofluorescent analysis of HCT 116 cells, wild-type (left, positive) or TBK1/NAK knockout (right, negative), using TBK1/NAK (E8I3G) Rabbit mAb (green). Actin filaments were labeled with Alexa Fluor® 555 Phalloidin #8953 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
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.
Flow cytometric analysis of THP-1 cells (green) using MAVS (D5A9E) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
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.
Flow cytometric analysis of TALL-1 cells (blue, negative) and Raji cells (green, positive) using TBK1/NAK (E8I3G) 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.
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To Purchase # 40415
Cat. # Size Qty. Price
40415T		 1 Kit  (9 x 20 microliters)

Bulk & Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
TBK1/NAK (E8I3G) Rabbit mAb 38066 20 µl
  • WB
  • IP
  • IF
  • F
 H M R 84 Rabbit IgG
MAVS (D5A9E) Rabbit mAb 24930 20 µl
  • WB
  • IP
  • IF
  • F
 H 75, 52 Rabbit IgG
TRAF2 (C192) Antibody 4724 20 µl
  • WB
  • IP
 H M Mk 53 Rabbit 
TRAF3 (E8H3B) Rabbit mAb 33640 20 µl
  • WB
  • IP
 H M R 62 Rabbit IgG
TRAF6 (E2K9D) Rabbit mAb 67591 20 µl
  • WB
  • IP
 H M R 60 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 (Ser386) (E7J8G) XP® Rabbit mAb 37829 20 µl
  • WB
  • IF
  • F
 H 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 MAVS Signalosome Antibody Sampler Kit provides an economical means of detecting the activation of signaling pathways in which MAVS plays a critical role. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the MAVS Signalosome Antibody Sampler Kit detects endogenous levels of its target protein. TRAF2 (C192) Antibody detects endogenous levels of total human TRAF2 protein. No cross-reactivity was detected with other family members at physiological conditions. Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb detects endogenous levels of TBK1 only when phosphorylated at Ser172. This antibody may cross-react with phospho-IKKε. Phospho-IRF-3 (Ser386) (E7J8G) XP® Rabbit mAb recognizes endogenous levels of IRF-3 protein only when phosphorylated at Ser386. Phospho-IRF-3 (Ser396) (D6O1M) Rabbit mAb recognizes endogenous levels of IRF-3 protein only when phosphorylated at Ser396.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Cys192 of human TRAF2. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with recombinant human IRF-3 protein and recombinant protein specific to a central region of mouse TRAF6, or synthetic phosphopeptides corresponding to residues surrounding Ser172 of human TBK1, Ser386 of human IRF-3, and Ser396 of human IRF-3 protein, or synthetic peptides corresponding to residues surrounding Leu202 of human MAVS and Ala327 of human TRAF3, or near the carboxy terminus of human TBK1/NAK protein.

Background

Recognition of conserved molecular structures of viruses by host pattern-recognition receptors (PRRs) initiates innate antiviral immune responses. Several families of PRRs have been demonstrated to sense different microbial components (1,2). The RIG-I-like receptor (RLR) family, including RIG-I, MDA5, and LGP2, are the primary PRRs to recognize viral RNAs (3,4). Upon binding to viral RNA, these receptors undergo conformation change, leading to their interaction with mitochondrial antiviral signaling protein (MAVS). MAVS subsequently forms large prion-like polymers and serves as a platform to recruit multiple components, including TRAF proteins and TBK1, to form the so-called MAVS signalosome. MAVS signalosome, in turn, activates the IRF-3 and NF-kB pathways, leading to the production of type I IFNs and pro-inflammatory cytokines (5-9).

Pathways

Explore pathways related to this product.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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