polyclonal antibody against myd88  (Shanghai Youke Equipment Instrument Co Ltd)

Journal: Frontiers in Immunology

Article Title: Dual roles of in situ generated HSP70 in antigen delivery and immunoregulation

doi: 10.3389/fimmu.2025.1638948

Figure Lengend Snippet: RFA induces TIRAP/MyD88 but not TRAM/TRIF association. (A–D) . Mice was subjected to RFA or Sham treatment or ID injection of 5 µg LPS. Skin was collected 6 h later and subjected to cryo-sectioning and PLA analysis of close association of MyD88 and TIRAP (A, B) as well as TRIF and TRAM (C, D) . Representative PLA images were shown in (A, C) and quantitative results were shown in (B, D) . Arrows point to PLA signals. E. Skin sections in RFA groups in the above studies were also stained with fluorescence-conjugated anti-MHC II antibodies. Z-stack pictures were captured and used to create 3D images. Representative 3D pictures showing the overlapping of RFA-induced TIRAP/MyD88 PLA signals with MHC II. Arrows point to overlapping signals (yellow). (F) Pie chart of PLA signals overlapped with MHC II (blue). Scale: 100 µm in (A, C, E) . One-way ANOVA with Newman-Keuls multiple comparison test was used to compare differences between groups in (B, D) . n=14–20 in (B) and n=6–8 in (D) . Over 50 PLA signals were explored by investigators in (F) . *, p<0.05; **, p<0.01; ***, p<0.001. Data are representative of three independent experiments with similar results.

Article Snippet: Goat polyclonal antibody against mouse/rat MyD88 (AF3109) was purchased from R & D Systems (Minneapolis, MN).

Techniques: Injection, Staining, Fluorescence, Comparison

Journal: Frontiers in Immunology

Article Title: Dual roles of in situ generated HSP70 in antigen delivery and immunoregulation

doi: 10.3389/fimmu.2025.1638948

Figure Lengend Snippet: HSP70 suppresses RFA-induced TLR4/IRAK/NFκB signaling. (A, B) WT and HSP70 KO mice were subjected to RFA or Sham treatment or ID injection of LPS or PBS. Skin was collected 6 h later in (A, B) IP and IB were conducted to evaluate TLR4/TIRAP binding (A) and IRAK4/IRAK1 binding in (B, C) . WT, HSP70 KO, MyD88 KO, TLR2 KO, and TLR4 KO mice were subjected to RFA or Sham treatment or ID injection of LPS. Skin was collected 2 h later. Cytosol and nuclear fractions were separated and nuclear fraction was analyzed by WB analysis to detect phosphorylated p65 using Lamin b1 as a loading control. (D) Skin IL-6 levels 6 h after RFA, Sham, or LPS treatment of lateral back skin of WT, HSP70 KO, TLR2 KO, TLR4 KO, and MyD88 KO mice. Two-way ANOVA with Fisher’s LSD test was used to compare differences between groups. n=4-6. *, p<0.05; **, p<0.01; ***, p<0.001. Original membrane pictures were shown in Supplementary Figure S9 . Data are representative of two independent experiments with similar results.

Article Snippet: Goat polyclonal antibody against mouse/rat MyD88 (AF3109) was purchased from R & D Systems (Minneapolis, MN).

Techniques: Injection, Binding Assay, Control, Membrane

Journal: PLOS Pathogens

Article Title: Arboviruses antagonize insect Toll antiviral immune signaling to facilitate the coexistence of viruses with their vectors

doi: 10.1371/journal.ppat.1012318

Figure Lengend Snippet: ( A) The accumulation levels of MyD88 in viruliferous S . furcifera were increased in MG132 treatment. (B) Schematic representation of the RING domain of SfREL. (C) Interaction between MyD88 and SfREL in Y2H assays. Transformants were plated on DDO or QDO with X-α-gal. DDO, SD/-Trp-Leu medium. QDO, SD/-Trp-Leu-His-Ade medium. (D) Interaction between MyD88 and SfREL in GST pull-down assay. GST-MyD88 was incubated with glutathione-Sepharose beads. SfREL-His was then added to the beads, followed by western blot assay to detect SfREL-His bound to GST-MyD88. (E) Interaction between MyD88 and SfREL-N or SfREL-C in Y2H assays. F-I, In vitro ubiquitination assay of GST-SfREL or GST-MyD88 in the presence of E1 and E2, as determined by western blot assays. (F and G) Ubiquitination of GST-SfREL or GST-SfREL (H61Y) was analyzed using ubiquitin (Ub) antibody. (H and I) Ubiquitination of GST-MyD88 by SfREL-HA or SfREL (H61Y)-HA was analyzed using Ub antibody. (J) Ubiquitination of GST-MyD88 by SfREL-HA was analyzed using Ub (k48) antibody. (K) Immunoprecipitation analysis of the ubiquitination of MyD88 in Sf9 cells expressed with MyD88-Flag, Ub-HA, together with SfREL-HA or GFP. The input proteins were analyzed using His-Tag, GST-Tag, or HA-Tag antibody.

Article Snippet: Rabbit polyclonal antibodies against SfREL, MyD88, Dorsal, and defensin of S . furcifera were prepared by GenScript Biotech Corporation in Nanjing, China.

Techniques: Pull Down Assay, Incubation, Western Blot, In Vitro, Ubiquitin Proteomics, Immunoprecipitation

Journal: PLOS Pathogens

Article Title: Arboviruses antagonize insect Toll antiviral immune signaling to facilitate the coexistence of viruses with their vectors

doi: 10.1371/journal.ppat.1012318

Figure Lengend Snippet: (A) Interaction between defensin and P10 in Y2H assays. Transformants were plated on either DDO or QDO. DDO, SD/-Trp-Leu medium. QDO, SD/-Trp-Leu-His-Ade medium. (B) Interaction between defensin and P10 in GST pull-down assay. GST-defensin was incubated with glutathione-Sepharose beads. P10-His was then added to the beads, followed by western blot assay to detect P10-His bound to GST-defensin. (C) Defensin-His and P10 singly expressed or co-expressed in Sf9 cells. Cells were respectively immunolabeled with His-Alexa Fluor 488 (green) or P10-Alexa Fluor 555 (red). The images were merged under a background of transmitted light. Panel i is the enlarged image of the boxed areas in left panel. Bars, 5 μm. (D and E) Immunoelectron microscopy showing the association of defensin with SRBSDV particles in virus-infected midgut. The intestines of nonviruliferous (D) and viruliferous (E) insects were immunolabeled with defensin antibody as the primary antibody, followed by treatment with 15-nm gold particle-conjugated IgG as the secondary antibody. Panel E-ii was the enlarged image of the boxed area in panel E-i. Red arrows indicate gold particles. V - , nonviruliferous; V + , viruliferous; Vi, virions. Bars, 100 nm. (F) Effects of membrane feeding of the mixture of purified defensin proteins and viral particles on the acquisition rates of SRBSDV in S . furcifera . The PBS buffer mixed with purified SRBSDV particles served as the control. Data are presented as means (± SD) of three independent biological replicates and each replicate contains 30 insects. **, P <0.01. ( G) Effects of defensin treatment through membrane feeding on the accumulation of SRBSDV P10 in 30 insects, as determined by western blot assay. (H) The intestines of insects microinjected with purified defensin or PBS mixed with purified SRBSDV particles were immunolabeled with P10-FITC (green). mg, midgut. Bars, 5 μm. (I) The average number of epithelial cells infected with SRBSDV in insects microinjected with defensin or PBS mixed with purified SRBSDV particles. Bars represent means ± SD from more than 20 individual cells. **, P <0.01. (J) Effects of the microinjected defensin on the accumulation of SRBSDV P10 in 30 insects, as determined by western blot assay. Insect GAPDH in G and J served as the reference of total proteins. The relative intensities of bands of P10 protein were determined using ImageJ.

Article Snippet: Rabbit polyclonal antibodies against SfREL, MyD88, Dorsal, and defensin of S . furcifera were prepared by GenScript Biotech Corporation in Nanjing, China.

Techniques: Pull Down Assay, Incubation, Western Blot, Immunolabeling, Immuno-Electron Microscopy, Virus, Infection, Membrane, Purification, Control

Journal: bioRxiv

Article Title: Ovine MyD88 have active role in host resistance against Haemonchus contortus infection

doi: 10.1101/2023.10.08.561398

Figure Lengend Snippet:

Article Snippet: Unlabelled goat anti-bovine polyclonal antibodies against MyD88 were obtained from Santa Cruz Biotechnology Inc, (CA, USA) and horseradish peroxidase-labeled rabbit anti-goat antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Amplification

Journal: bioRxiv

Article Title: Ovine MyD88 have active role in host resistance against Haemonchus contortus infection

doi: 10.1101/2023.10.08.561398

Figure Lengend Snippet: TIR domain for MyD88 from amino acid position 159 to 293aa.

Article Snippet: Unlabelled goat anti-bovine polyclonal antibodies against MyD88 were obtained from Santa Cruz Biotechnology Inc, (CA, USA) and horseradish peroxidase-labeled rabbit anti-goat antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: bioRxiv

Article Title: Ovine MyD88 have active role in host resistance against Haemonchus contortus infection

doi: 10.1101/2023.10.08.561398

Figure Lengend Snippet: STRING Network analysis for MYD88 with other genes in sheep

Article Snippet: Unlabelled goat anti-bovine polyclonal antibodies against MyD88 were obtained from Santa Cruz Biotechnology Inc, (CA, USA) and horseradish peroxidase-labeled rabbit anti-goat antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: bioRxiv

Article Title: Ovine MyD88 have active role in host resistance against Haemonchus contortus infection

doi: 10.1101/2023.10.08.561398

Figure Lengend Snippet: KEGG pathway analysis depicted the role of MyD88 through TLR signalling pathway

Article Snippet: Unlabelled goat anti-bovine polyclonal antibodies against MyD88 were obtained from Santa Cruz Biotechnology Inc, (CA, USA) and horseradish peroxidase-labeled rabbit anti-goat antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: bioRxiv

Article Title: Ovine MyD88 have active role in host resistance against Haemonchus contortus infection

doi: 10.1101/2023.10.08.561398

Figure Lengend Snippet: KEGG pathway analysis depicted the role of MyD88 through MAPK signalling pathway

Article Snippet: Unlabelled goat anti-bovine polyclonal antibodies against MyD88 were obtained from Santa Cruz Biotechnology Inc, (CA, USA) and horseradish peroxidase-labeled rabbit anti-goat antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: bioRxiv

Article Title: Ovine MyD88 have active role in host resistance against Haemonchus contortus infection

doi: 10.1101/2023.10.08.561398

Figure Lengend Snippet: KEGG pathway analysis depicted the role of MyD88 through NF kappa signalling pathway

Article Snippet: Unlabelled goat anti-bovine polyclonal antibodies against MyD88 were obtained from Santa Cruz Biotechnology Inc, (CA, USA) and horseradish peroxidase-labeled rabbit anti-goat antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: bioRxiv

Article Title: Ovine MyD88 have active role in host resistance against Haemonchus contortus infection

doi: 10.1101/2023.10.08.561398

Figure Lengend Snippet: KEGG pathway analysis depicted the role of MyD88 through NOD like receptor pathway

Article Snippet: Unlabelled goat anti-bovine polyclonal antibodies against MyD88 were obtained from Santa Cruz Biotechnology Inc, (CA, USA) and horseradish peroxidase-labeled rabbit anti-goat antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: bioRxiv

Article Title: Ovine MyD88 have active role in host resistance against Haemonchus contortus infection

doi: 10.1101/2023.10.08.561398

Figure Lengend Snippet: KEGG pathway analysis depicted the role of MyD88 against toxoplasmosis

Article Snippet: Unlabelled goat anti-bovine polyclonal antibodies against MyD88 were obtained from Santa Cruz Biotechnology Inc, (CA, USA) and horseradish peroxidase-labeled rabbit anti-goat antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: bioRxiv

Article Title: Ovine MyD88 have active role in host resistance against Haemonchus contortus infection

doi: 10.1101/2023.10.08.561398

Figure Lengend Snippet: KEGG pathway analysis depicted the role of MyD88 against African Trypanosomiasis

Article Snippet: Unlabelled goat anti-bovine polyclonal antibodies against MyD88 were obtained from Santa Cruz Biotechnology Inc, (CA, USA) and horseradish peroxidase-labeled rabbit anti-goat antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: bioRxiv

Article Title: Ovine MyD88 have active role in host resistance against Haemonchus contortus infection

doi: 10.1101/2023.10.08.561398

Figure Lengend Snippet: KEGG pathway analysis depicted the role of MyD88 against Chagas disease

Article Snippet: Unlabelled goat anti-bovine polyclonal antibodies against MyD88 were obtained from Santa Cruz Biotechnology Inc, (CA, USA) and horseradish peroxidase-labeled rabbit anti-goat antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques:

Journal: Scientific Reports

Article Title: miR-628, a microRNA that is induced by Toll-like receptor stimulation, regulates porcine innate immune responses

doi: 10.1038/srep12226

Figure Lengend Snippet: Shown is a sequence alignment of miR-628 and its target sites in 3′UTR of MyD88. Also shown is an analysis of expression of MyD88-UTR luciferase reporters in the presence of miR-628/inhibitor or the irrelevant control (IC). Filled bars correspond to reporter constructs with wild-type miR-628 targeting sites, and open bars correspond to constructs with 6-nt substitutions disrupting base-pairing with the “seed region” of miR-628. (* means P<0.05; **means P<0.01).

Article Snippet: The separated proteins were blotted on to a PVDF membrane and probed with a rabbit polyclonal antibody against amino acids at the C-terminus of MyD88 and NF-κB of porcine origin (Santa Cruz Biotechnol, Inc.).

Techniques: Sequencing, Expressing, Luciferase, Control, Construct

Journal: Scientific Reports

Article Title: miR-628, a microRNA that is induced by Toll-like receptor stimulation, regulates porcine innate immune responses

doi: 10.1038/srep12226

Figure Lengend Snippet: ( A ) Western-blot analysis of MyD88 protein in porcine monocytes. ( B ) Western-blot analysis of nucleus NF-κB protein in porcine monocytes. CON, non-stimulated cells; CONM, CON + mimics; CONI, CON + inhibitors; CONMI, CON + mimics + inhibitors; LPS, LPS-stimulated cells; LPSM, LPS+ mimics; LPSI, LPS- + inhibitors; LPSMI, LPS + mimics + inhibitors. (* means P<0.05; **means P<0.01).

Article Snippet: The separated proteins were blotted on to a PVDF membrane and probed with a rabbit polyclonal antibody against amino acids at the C-terminus of MyD88 and NF-κB of porcine origin (Santa Cruz Biotechnol, Inc.).

Techniques: Western Blot