Log In Sign up
The largest database of trusted experimental protocols

Poly da dt

Manufactured by InvivoGen
Visit Supplier
Sourced in United States, Hong Kong, France

Poly(dA:dT) is a synthetic double-stranded DNA molecule composed of alternating adenine (dA) and thymine (dT) nucleotides. It is commonly used as a laboratory tool to study various biological processes and as a control in molecular biology experiments.

Automatically generated - may contain errors

Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (15 mentions) Poly 1 c, by InvivoGen (13 mentions) Nigericin, by InvivoGen (13 mentions) Pam3csk4, by InvivoGen (10 mentions) Adenosine triphosphate (atp), by Merck Group (9 mentions) Lipopolysaccharides (lps), by InvivoGen (8 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (8 mentions) Opti mem, by Thermo Fisher Scientific (7 mentions) Poly 1 c lmw, by InvivoGen (7 mentions) Nigericin, by Merck Group (6 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions) 2 3 cgamp, by InvivoGen (5 mentions) Lipopolysaccharides (lps), by Merck Group (5 mentions) Flagellin, by InvivoGen (5 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (4 mentions) Il 1β, by R&D Systems (3 mentions) Mg132, by Merck Group (3 mentions) Ifn β, by R&D Systems (3 mentions) Poly 1 c hmw, by InvivoGen (3 mentions)

Spelling variants of this product

Poly(dA:dT)/LyoVec (15 citations) Poly(dA:dT; tlrl-patn) (4 citations) Poly(dA:dT) naked (3 citations) Poly(dA:dT)-Lyovec complexes (2 citations) Polydeoxyadenylic-deoxythymidylic acid sodium salt (poly(dA:dT)) (2 citations)

90 protocols using poly da dt

1

Culture and Infection of Bone Marrow-Derived Macrophages

Check if the same lab product or an alternative is used in the 5 most similar protocols
Bone marrow-derived macrophages (BMDMs) were cultured for 6 days in macrophage culture medium, that is DMEM supplemented with 10% FBS, 30% L929 conditioned media and 1% penicillin and streptomycin, and seeded at a concentration of 1 × 106 cells onto 12-well plates. Next day, BMDMs were incubated in antibiotic-free media. F. novicida was added to BMDMs at an indicated MOI for 20 hours. 50 μg/ml gentamicin was added 4 hours after the infection. The MCMV strain (K181) was obtained from Edward S. Mocarski (Emory University School of Medicine) (32 (link)). MCMV was added at an MOI of 10 for 10 hours. For poly(dA:dT) transfection, 1 μg of poly(dA:dT) (InvivoGen) was resuspended in PBS, mixed with 0.3 μl of Xfect polymer in Xfect reaction buffer (Clontech Laboratories) and incubated for 10 minutes. DNA complexes were added to BMDMs in Opti-MEM and incubated for 3 hours.
Zhu Q., Zheng M., Balakrishnan A., Karki R, & Kanneganti T.D. (2018). Gasdermin D promotes AIM2 inflammasome activation and is required for host protection against Francisella novicida. Journal of immunology (Baltimore, Md. : 1950), 201(12), 3662-3668.
+ Open protocol
+ Expand
2

Inflammasome Activation in BMDMs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immortalized BMDMs (seeded at 250 000 cells per ml in 96-well plates) were primed for 4 h with 100 ng ml−1 LPS O55:B5 (Invivogen). The NLRP3 inflammasome was triggered by addition of 5 mM extracellular ATP (Sigma-Aldrich) for 60 min. The AIM2 inflammasome was triggered by transfection of 1 μg ml−1 of the synthetic DNA analogue poly(dA:dT) (Invivogen) using Lipofectamine 2000 (Invitrogen), according to the manufacturer's protocol, in OptiMEM (Gibco) for 3 h. The NLRC4 inflammasome was triggered by infection of the cells with S. enterica serovar Typhimurium SL1344 at a multiplicity of infection of 10. The infection was synchronized by centrifugation and continued for 60 min. The pyrin inflammasome was triggered by addition of 1 μg ml−1 (final concentration) of C. difficile toxin B (CdtB, Enzo Biotech) for 2.5 h.
Dick M.S., Sborgi L., Rühl S., Hiller S, & Broz P. (2016). ASC filament formation serves as a signal amplification mechanism for inflammasomes. Nature Communications, 7, 11929.
+ Open protocol
+ Expand
3

Inflammasome Activation Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
LPS (O111:B4, L2630), ATP (A7699), disuccinimidyl suberate (S1885), nigericin (N7143), and phorbol myristate acetate (PMA, P1585) were from Sigma-Aldrich; alum (77161) and the c-Myc tag (MA1-980) were from Thermo Fisher Scientific; and poly(dA:dT) was from InvivoGen. The following antibodies were used: anti-Flag (F1804) (Sigma); anti-mouse caspase-1 p20 (AG-20B-0042) and anti-mouse NLRP3 (AG-20B-0014) (AdipoGen); anti-human caspase-1 (ab108362) and anti-BRCC36 (ab108411) (Abcam); anti-mouse IL-1β (AF-401-NA) (R&D); anti-HA-tag (3724) (Cell Signaling Technology); and anti-VDR (sc-13133), anti-ASC (sc-514414), and anti-β-actin (sc-47778) (Santa Cruz). The Mouse TNF ELISA Kit (558534), Mouse IL-1β ELISA Kit (559603), and TMB Substrate Reagent Set (555214) were from BD Biosciences. The mouse IL-18 ELISA set (EMC06) was from ExCell Biotech (Table 1).
Rao Z., Chen X., Wu J., Xiao M., Zhang J., Wang B., Fang L., Zhang H., Wang X., Yang S, & Chen Y. (2019). Vitamin D Receptor Inhibits NLRP3 Activation by Impeding Its BRCC3-Mediated Deubiquitination. Frontiers in Immunology, 10, 2783.
+ Open protocol
+ Expand
4

Murine cell differentiation protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
C57BL/6 male mice (6-8 weeks of age) and Ifnar1 −/− mice were from the Jackson Laboratory. All mice experiments were performed in accordance with guidelines from the University of California, Los Angeles, Institutional Animal Care and Use Committee. HEK293T and RAW264.7 cells were purchased from ATCC and cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. RXRαfl/fl (WT) and LysM-Cre +/RXRαfl/fl (Rxra−/−) mice bone marrows were overnight shipped from Dr Mercedes Ricote's Lab (Spain)13 (link). WT, Ifnar1 −/− and Rxra −/− BMMs were differentiated as described previously45 (link). WT and Rxra −/− F9 embryocarcinoma cells, as well as RXR-specific agonists AGN194204 and LG268 were obtained from Dr Peter Tontonoz's Lab (University of California, Los Angeles). HX531 was obtained from Dr Hiroyuki Kagechika's lab (Tokyo Medical and Dental University). 9cRA and 13cRA were purchased from Sigma-Aldrich. All compounds for in vitro treatment were solubilized in DMSO. PolyI:C and polydA:dT were from InvivoGen. Antibody against α-tubulin was from Sigma-Aldrich. Anti-VSV-G (P5D4) and anti-Oct1 (C-21) antibodies were from Santa Cruz Biotechnology. Anti-RXRα (D6G10), anti-β-Actin (13E5) and anti-β-Catenin (D10A8) were from Cell Signaling Technology. Anti-GAPDH (GT239) was from GeneTex.
Ma F., Liu S.Y., Razani B., Arora N., Li B., Kagechika H., Tontonoz P., Núñez V., Ricote M, & Cheng G. (2014). Retinoid X receptor α attenuates host antiviral response by suppressing type I interferon. Nature communications, 5, 5494.
+ Open protocol
+ Expand
5

Viral Infection and Immune Response Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
HSV-1 virus stock (VR-733) was purchased from ATCC (Manassas, VA). PRR agonists CpG (ODN2395), CL264 (cat#: tlrl-c264e), LPS (cat#: tlrl-b5LPS), Pam3CSK4 (cat#: tlrl-pms), PolydA:dT (cat#: tlrl-patc), PolyI:C-LMW (cat#:tlrl-picwlv), polyI:C-TLR3 agonist (cat#: tlrl-picw), and SS40 (cat#: tlrl-lrna40) were purchased from InvivoGen (San Diego, CA). Recombinant human TNFα (cat# 210-TA-020/CF), IL-1β (cat# 201-LB-005), IFNγ, IL-4, IL-22, GM-CSF, monoclonal mouse IgG1 Clone#11711 (Cat# MAB002), human IL-1β/IL-1F2 antibody (MAB601), human TNFαR1/TNFRSF1A antibody (MAB625), and anti-human IFNγR1 antibody (MAB6731) were bought from R&D systems, Inc. (Minneapolis, MN). Recombinant human IFNα (cat#:11101-1) and mouse anti-human IFNα (cat#: 21112-1) were purchased from PBL Biomedical Laboratories (Piscataway, NJ). Myc-DDK1-tagged RELA (RC220780), Myc-DDK1-tagged NFKB1 (RC208384), Myc-DDK1-tagged IRF7 (RC217934), Myc-DDK1-tagged ANKRD1 (RC205609) and Myc-DDK1-tagged STING were purchased from OriGene (Rockville, MD). Dr. Hong-Bing Shu (Wuhan University, China) kindly provided pCMV-flag-IRF3 and pCMV-flag-MyD88. PRK-neo-HA-ANKRD1 was generated in our lab by insertion of an encoding cDNA fragment in frame into PRK-neo-HA vector (a kind gift from Dr. Hong-Bing Shu).
Bin L., Li X., Richers B., Streib J.E., Hu J.W., Taylor P, & Leung D.Y. (2018). ANKRD1 regulates innate immune responses against herpes simplex virus 1: A potential role in eczema herpeticum. The Journal of allergy and clinical immunology, 141(6), 2085-2093.e1.
+ Open protocol
+ Expand
6

SARS-CoV-2 and MERS-CoV Protein Expression

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Vero E6, Huh-7, 293T, and HeLa cells were grown in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), and Calu-3 2B4 cells (Kent Tseng, University of Texas Medical Branch) were grown in MEM supplemented with 20% FBS as previously described (41 (link)). Codon-optimized sMacro (nucleotides 3262 to 3783 of SARS-CoV MA15) and MERS-CoV ORF4a were synthesized and cloned into pUC57 (GenScript). The sMacro and GFP sequences were PCR amplified and ligated into a linearized pcDNA3 plasmid using In-Fusion (Invitrogen) cloning. The ORF4a sequence was PCR amplified and then restriction digested and ligated into the pLKO plasmid. The resulting constructs were confirmed by restriction digestion, PCR, and direct sequencing. Human IFN-α (B/D) and IFN-β were purchased from PBL (Piscataway, NJ). High-molecular-weight poly(I-C) and poly(dA-dT) were purchased from InvivoGen (San Diego, CA). Cells were transfected with either Polyjet (Amgen, Thousand Oaks, CA) or Lipofectamine 2000 (Fisher Scientific, Waltham, MA) per the instructions of the manufacturers.
Fehr A.R., Channappanavar R., Jankevicius G., Fett C., Zhao J., Athmer J., Meyerholz D.K., Ahel I, & Perlman S. (2016). The Conserved Coronavirus Macrodomain Promotes Virulence and Suppresses the Innate Immune Response during Severe Acute Respiratory Syndrome Coronavirus Infection. mBio, 7(6), e01721-16.
+ Open protocol
+ Expand
7

Virulent, Recombinant, and Attenuated DPV Strains

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The virulent strain of DPV, CHv, was isolated and characterized by our lab (32 (link)). The recombinant virulent strain of DPV, BAC-CHv-EGFP (CHv-GFP), was constructed by our research center (33 (link)). The attenuated vaccine strain of DPV, CHa, was retrieved from storage at our research center. The Duck Tambusu virus (DTMUV) was stored at our research center. Each DPV and DTMUV strain was propagated on DEFs. The Rabbit anti-human MAP2 and -human GFAP polyclonal antibody and the goat anti-human β-actin monoclonal antibody were purchased from Abclonal (Wuhan, China). Mouse anti-duck CD80 (Gene ID:101796029) and CD86 (Gene ID:101794331) antibodies were generated by our research center. Ruxolitinib, poly(dA:dT), and poly(I:C) were purchased from Invivogen (Hong Kong, China).
Tian B., Cai D., He T., Deng L., Wu L., Wang M., Jia R., Zhu D., Liu M., Yang Q., Wu Y., Zhao X., Chen S., Zhang S., Huang J., Ou X., Mao S., Yu Y., Zhang L., Liu Y, & Cheng A. (2020). Isolation and Selection of Duck Primary Cells as Pathogenic and Innate Immunologic Cell Models for Duck Plague Virus. Frontiers in Immunology, 10, 3131.
+ Open protocol
+ Expand
8

Inflammasome Activation in 293T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
293T cells (ATCC source) (1 × 106 /mL) were co-transfected with 0.3 μg, 0.1 μg, and 0.2 μg of expression plasmids encoding human Flag-tagged pro-IL-1β, Flag-tagged pro-casp1, and Myc-tagged ASC, respectively, and with 0.3 μg of plasmid for co-expression of GFP with NLRP3, NLRP1, NLRC4, or AIM2. The expression of these inflammasome components was confirmed by western blotting with a Myc-tag antibody (CST, 2272S, USA) and a FLAG-tag antibody (MBL, M185-3L, USA) and by examination of GFP expression with a fluorescence microscope (Nikon 80I; Tokyo, Japan).
At 24 h post-transfection, cells were infected with S. suis strain SC-19 for 2 h or transfected with poly(dA:dT) (Invivogen, tlrl-patn, USA) for 12 h. Then, cell supernatants were collected for the western blot assay with antibodies against casp1 (Cell Signaling, 3866S, USA) and IL-1β (Proteintech, 16806-1-AP, USA) and for determination of IL-1β (eBioscience, 88-7261-88, USA).
Lin L., Xu L., Lv W., Han L., Xiang Y., Fu L., Jin M., Zhou R., Chen H, & Zhang A. (2019). An NLRP3 inflammasome-triggered cytokine storm contributes to Streptococcal toxic shock-like syndrome (STSLS). PLoS Pathogens, 15(6), e1007795.
+ Open protocol
+ Expand
9

Deciphering Innate Immune Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Rabbit antibodies against human p-TBK1 Ser172 (catalog no. 5483), TBK1 (catalog no. 3504), IRF3 (catalog no. 11904), p-STING Ser366 (catalog no. 85735), and STING (catalog no. 13647) were obtained from Cell Signaling Technology. Rabbit antibodies against human p-IRF3 Ser386 (ab76493), MAVS (ab31334), and green fluorescent protein (GFP) (ab290) were purchased from Abcam. Rabbit antibody against HA was purchased from HUAXINGBIO (HX1820). Mouse antibodies against laminB1 (sc-365962) were purchased from Santa Cruz Biotechnology. Mouse anti-FLAG antibody (M2) (F1804), M2-conjugated agarose (A2220), mouse anti-HA antibody (H3663), and anti-HA–conjugated agarose (A2095) were purchased from Sigma-Aldrich. Rabbit anti-PPM1G antibody (A300-881A) was purchased from Bethyl Laboratories. Poly(I:C) and poly(dA:dT) were purchased from InvivoGen (tlrl-pic, tlrl-patn-1). Mouse anti-ORF33 antibody was provided by F. Zhu (Florida State University, USA). Mouse anti-K8/KbZIP and anti-ORF45 antibodies were provided by Y. Yuan (University of Pennsylvania, USA). Human IFNβ ELISA Kit was purchased from R&D Systems (DIFNB0). Duolink In Situ–Fluorescence kit for PLA was purchased from Sigma-Aldrich (DUO92008, DUO92002, and DUO92004).
Yu K., Tian H, & Deng H. (2020). PPM1G restricts innate immune signaling mediated by STING and MAVS and is hijacked by KSHV for immune evasion. Science Advances, 6(47), eabd0276.
+ Open protocol
+ Expand
10

Bone Marrow Macrophage Stimulation Assay

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Bone marrow–derived macrophages were prepared as described previously (21 (link)). Cells were stimulated with LPS (500 ng/mL; InvivoGen), Pam3CSK4 (1 μg/mL), gardiquimod (1 μg/mL; InvivoGen), Poly(I:C) (10 μg/mL; InvivoGen), or IFNβ (400 U/mL; PBL Assay Science). For transfection of DNA or 2´3´-cyclic guanosine monophosphate–adenosine monophosphate (cGAMP), 0.25 μg of poly(dA:dT) (InvivoGen) or 1 μg of 2´3´-cGAMP (InvivoGen) was mixed with 0.3 mL of Xfect polymer in Xfect reaction buffer (Clontech Laboratories) for 10 min and then added to BMDMs in Opti-MEM (ThermoFisher Scientific).
Zhu Q, & Kannegant T.D. (2017). Distinct regulatory mechanisms control proinflammatory cytokines IL-18 and IL-1β. Journal of immunology (Baltimore, Md. : 1950), 198(11), 4210-4215.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!