Journal: Cell reports

Article Title: Unannotated microprotein EMBOW regulates the interactome and chromatin and mitotic functions of WDR5

doi: 10.1016/j.celrep.2023.113145

Figure Lengend Snippet:

Article Snippet: MYC Epitope Tag Antibody Rabbit Polyclonal , Rockland , Cat#600-401-381; RRID:AB_217927.

Techniques: Control, Recombinant, Affinity Purification, Virus, Plasmid Preparation, Quantitative Proteomics, Over Expression, Knock-Out, Software

Journal: Nature Immunology

Article Title: Cross-presentation of dead cell-associated antigens shapes the neoantigenic landscape of tumor immunity

doi: 10.1038/s41590-025-02354-w

Figure Lengend Snippet: a , Schematic illustration of the two OVA fusion constructs, LA-OVA (top) and the control mutLA-OVA (bottom). b , Western blot analysis of lysates from transfected and untransfected HeLa cells probed with anti-His (top), anti-OVA (middle) and anti-β-actin (bottom) mAbs. c , Confocal analysis of HeLa cells expressing LA-OVA (left figure, top panels) or mutLA-OVA (left figure, bottom panels). Intensity profiles for OVA (AF647) and phalloidin (AF488) channels across 10 μm (white line indicated in the confocal images) are shown on the right. Scale bars shown in confocal images (left) correspond to a size of 5μm. d - e , Flow cytometry analysis of untransfected and transfected HeLa cells before (live) and after UV treatment (UV). Overlay histograms represent intracellular staining for OVA ( d , top panels) and e ). RFP fluorescence is shown for LA-OVA and mutLA-OVA cells ( d , bottom panels). Numbers in the histogram overlay represent mean fluorescence intensity. Data are representative of 3 independent experiments.

Article Snippet: HRP-conjugated goat-anti-rabbit and HRP-conjugated mouse-anti-His-Tag antibodies were from Southern Biotech.

Techniques: Construct, Control, Western Blot, Transfection, Expressing, Flow Cytometry, Staining, Fluorescence

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Cutting edge: CD4 is the receptor for the tick saliva immunosuppressor, Salp15.

doi: 10.4049/jimmunol.177.10.6579

Figure Lengend Snippet: FIGURE 1. Salp15 binds to CD4. A, Left panels, Analysis of His-tagged Salp15-488 binding to CD4 and CD8 T cells by confocal microscopy compared with unstained CD4 T cells (control). The panels on the right show the same field under brightfield microscopy. B, Murine primary CD4 T cells lysates containing His-tagged Salp15 were immunoprecipitated (IP) using anti-His, anti-CD3, anti-CD28, anti-CD4, anti-TCR, or IgG (control). The immunoprecipitate was immunoblotted (IB) using anti-His, anti-CD4, anti-CD3, anti-TCR, and anti-CD28 Abs. C, Colocalization of anti-CD4 staining and His-tagged Salp15-488 binding on naive and stimulated CD4 T cells is indicated by the yellow color in the merged confocal micrograph. D, Flow cytometric analysis of CD4 expression using PE-Cy5-labeled anti-CD4 (CD4PE-Cy5) (upper panel) and His-tagged Salp15-488 binding (lower panel) in 3T3 (red) and 3T3-CD4 (blue) cells. E, HeLa and HeLa-CD4 cell lysates containing His-tagged Salp15 were immunoprecipitated using anti-His Ab. The immunoprecipitates were subjected to immunoblotting using anti-CD4 and anti-His Abs. The reciprocal immunoprecipitation from a HeLa-CD4 cell lysate was performed using anti-CD4 or IgG followed by immu- noblotting with anti-CD4 or anti-His Abs. F, Immunoprecipitation from HeLa-CD4 cell lysate containing either His-tagged TR-Salp13 (control) or His-tagged Salp15 using an anti-His Ab followed by immunoblotting with anti-CD4 or anti-His Abs.

Article Snippet: The polyclonal anti-His-HRP Ab was obtained from Novus Biologicals.

Techniques: Binding Assay, Confocal Microscopy, Control, Microscopy, Immunoprecipitation, Staining, Expressing, Labeling, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Cutting edge: CD4 is the receptor for the tick saliva immunosuppressor, Salp15.

doi: 10.4049/jimmunol.177.10.6579

Figure Lengend Snippet: FIGURE 2. Salp15 binds to the extracellular domains of CD4. A, Unlabeled Salp15 but not lysozyme pretreatment blocks His-tagged Salp15-488 binding to HeLa-CD4 cells. Control represents HeLa-CD4 cells in the absence of His-tagged Salp15-488. Right panel, The brightfield of the image on the left. B, Preincubation of HeLa-CD4 cells with a polyclonal anti-CD4 Ab abolishes His-tagged Salp15-488 binding compared with control IgG pretreatment. C, His-tagged Salp15 (0.4 M) was incubated with increasing amounts of immo- bilized sCD4 (D1D2, f) or lysozyme (Œ) in a microtiter assay showing satu- rable binding. The results are expressed as mean SE of three independent experiments. D, Elution profiles of sCD4 (D1–D4) (blue), His-tagged Salp15 (pink), and His-tagged Salp15-sCD4 (red) from Superdex-200 gel filtration columns (left panel). The Gaussian deconvolution of His-tagged Salp15-sCD4 and sCD4 peaks is shown on the right panel. The results shown are representa- tive of three to five individual experiments performed.

Article Snippet: The polyclonal anti-His-HRP Ab was obtained from Novus Biologicals.

Techniques: Binding Assay, Control, Incubation, Filtration

Journal: Vaccines

Article Title: Evaluation of Immunogenicity and Efficacy of Fasciola hepatica Tetraspanin 2 (TSP2) Fused to E. coli Heat-Labile Enterotoxin B Subunit LTB Adjuvant Following Intranasal Vaccination of Cattle

doi: 10.3390/vaccines9111213

Figure Lengend Snippet: Characterisation of LTB oligomerisation. Both proteins rLTB-FhTSP2 ( a , c ) and rLTB ( b , d ) were loaded at 6 µg and subject to boiling at 95 °C for 10 min. Boiled (lane 1) and non-boiled (lane 2) proteins were analysied by SDS-PAGE and visualized by Coomassie staining ( a , b ) and Western blotting ( c , d ) using anti-His tag antibodies.

Article Snippet: After washing five times with 1× PBS-T, plates were incubated for 1 h at RT with 100 μL/well of mouse anti-His tag HRP conjugate antibodies (R&D Systems) diluted 1:10,000 in blocking buffer C. Plates were washed five times with 1× PBS-T with the reactions developed and stopped and the plate OD read as described above.

Techniques: SDS Page, Staining, Western Blot

Journal: Vaccines

Article Title: Evaluation of Immunogenicity and Efficacy of Fasciola hepatica Tetraspanin 2 (TSP2) Fused to E. coli Heat-Labile Enterotoxin B Subunit LTB Adjuvant Following Intranasal Vaccination of Cattle

doi: 10.3390/vaccines9111213

Figure Lengend Snippet: Characterisation of pentamer formation of the rLTB-FhTSP2 fusion complex via glycan receptor binding. Plates were coated with 3 μg/mL of GM1 ganglioside and blocked with 3% bovine serum albumin. Potential binding proteins (rEcxB and rLTB-FhTSP2) and negative control (rFhTSP2) were incubated in triplicate at three dilution factors (3, 0.3, and 0.03 μg/mL). Binding was detected by mouse anti-His tag HRP conjugated antibodies. All results are displayed as the mean of the adjusted optical density after subtraction of non-GM1 coated wells.

Article Snippet: After washing five times with 1× PBS-T, plates were incubated for 1 h at RT with 100 μL/well of mouse anti-His tag HRP conjugate antibodies (R&D Systems) diluted 1:10,000 in blocking buffer C. Plates were washed five times with 1× PBS-T with the reactions developed and stopped and the plate OD read as described above.

Techniques: Glycoproteomics, Binding Assay, Negative Control, Incubation

Journal: Vaccines

Article Title: Evaluation of Immunogenicity and Efficacy of Fasciola hepatica Tetraspanin 2 (TSP2) Fused to E. coli Heat-Labile Enterotoxin B Subunit LTB Adjuvant Following Intranasal Vaccination of Cattle

doi: 10.3390/vaccines9111213

Figure Lengend Snippet: Characterisation of the antigenicity of the rLTB-FhTSP2 fusion complex. The reactivity of rFhTSP2, rLTB-FhTSP2, and rLTB was analysed using diluted bovine anti-rFhTSP2 sera (1:100, 1:200, 1:400, and 1:800) and a anti-bovine IgG HRP secondary antibody. Antigens were coated at an equal molar ratio of the FhTSP2 protein sequence.

Article Snippet: After washing five times with 1× PBS-T, plates were incubated for 1 h at RT with 100 μL/well of mouse anti-His tag HRP conjugate antibodies (R&D Systems) diluted 1:10,000 in blocking buffer C. Plates were washed five times with 1× PBS-T with the reactions developed and stopped and the plate OD read as described above.

Techniques: Sequencing

Journal: Vaccines

Article Title: Evaluation of Immunogenicity and Efficacy of Fasciola hepatica Tetraspanin 2 (TSP2) Fused to E. coli Heat-Labile Enterotoxin B Subunit LTB Adjuvant Following Intranasal Vaccination of Cattle

doi: 10.3390/vaccines9111213

Figure Lengend Snippet: Bovine humoral response towards rFhTSP2 over the course of the trial. ( a ) Serum IgG, ( b ) serum IgA, and ( c ) saliva IgA responses from Day 0 (pre-vaccination), Day 42 (post-vaccination), Day 84 (post-challenge), and Day 125 (end of trial) from Group 1 (G1: dark grey) subcutaneously vaccinated with PBS; Group 2 (G2: red) intranasally vaccinated with rLTB-FhTSP2; and Group 3 (G3: blue) subcutaneously vaccinated with rFhTSP2. Each animal within a group is represented by a different shape. The reactivity of 5 μg/mL of rFhTSP2 was analysed using serum (diluted 1:50) or saliva (diluted 1:8) and an anti-bovine IgG HRP secondary antibody diluted 1:5000 or a anti-bovine IgA HRP antibody diluted 1:2000. The mean OD of each group is represented by a solid line. Significant differences ( p < 0.05) of the mean are denoted by asterisks (*).

Article Snippet: After washing five times with 1× PBS-T, plates were incubated for 1 h at RT with 100 μL/well of mouse anti-His tag HRP conjugate antibodies (R&D Systems) diluted 1:10,000 in blocking buffer C. Plates were washed five times with 1× PBS-T with the reactions developed and stopped and the plate OD read as described above.

Techniques: