recombinant human argonaute 2 eif2c2 protein  (Sino Biological)

Journal: Journal of Nanobiotechnology

Article Title: Treatment of breast cancer with autophagy inhibitory microRNAs carried by AGO2-conjugated nanoparticles

doi: 10.1186/s12951-020-00615-4

Figure Lengend Snippet: Physical characterization and composition of SPION nanoparticles

Article Snippet: Human AGO2 (His-tag) (EIF2C2) recombinant protein was purchased from Sino Biological Inc. (China).

Techniques:

Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: A Novel Method to Silence Genes in Cryptosporidium

doi: 10.1007/978-1-4939-9748-0_11

Figure Lengend Snippet: Silencing genes in Cryptosporidium. (a) Assembling: hAgo2 protein is loaded with Cryptosporidium ssRNA complementary to mRNA target. (b) Encapsulation: hAgo2–ssRNA complex is encapsulated within liposomes (protein transfection reagent). (c) Transfection: Cryptosporidium oocysts are transfected with complexes. (d) Silencing: hAgo2–ssRNA binds to mRNA target, translation is blocked, and mRNA target is sliced by hAgo2, then expression of target is reduced

Article Snippet: Recombinant protein human Argonaute 2 (hAgo2) from human (Sino Biologicals, North Wales, PA).

Techniques: Transfection, Expressing

Journal: Cellular and Molecular Life Sciences

Article Title: The RNA–RNA base pairing potential of human Dicer and Ago2 proteins

doi: 10.1007/s00018-019-03344-6

Figure Lengend Snippet: hAgo2 displays limited RNA-annealing activity. a – c Native PAGE gels showing the results of annealing reactions involving a R21 and Mod18, b R21 and Mod23, and c R21 and Mod33. Reaction mixtures were incubated with increasing amounts of hAgo2 or with no protein (“hyphen”). Graphs showing representative annealing reactions obtained by densitometric quantification of the autoradiograms (right panel). Horizontal dashed lines are drawn for the values obtained for control experiments with no protein (baselines). d – f Time-dependent annealing assays involving d R21 and Mod18, e R21 and Mod23, and f R21 and Mod33. Graphic presentation of the results obtained from three independent annealing assays involving hAgo2 (in orange) or no protein (in blue). The x -axis represents the incubation time expressed in minutes, and the y -axis represents the percentage content of the R21-Mod duplex fraction

Article Snippet: Recombinant human Ago2 (hAgo2) protein was purchased from Active Motif, Giardia intestinalis endoribonuclease Dicer-like recombinant protein (GiDicer) was from MyBiosource, and recombinant human Dicer (hDicer) was produced in our laboratory.

Techniques: Activity Assay, Clear Native PAGE, Incubation, Control

Journal: Cellular and Molecular Life Sciences

Article Title: The RNA–RNA base pairing potential of human Dicer and Ago2 proteins

doi: 10.1007/s00018-019-03344-6

Figure Lengend Snippet: hDicer accelerates the annealing of miRNA to its target site present within the DICER1 transcript. a A scheme representing the base pairing of miR-103a-3p with its target site within the DICER1 transcript. b – d Native PAGE gels showing the results of annealing reactions involving 5 nM of 3′- 32 P-labeled Ex21 and 50 nM of either b miRNA-103a-3p, c miRNA-103a-5p or d miRNA-103a duplex. Reaction mixtures were incubated with either 25 nM hDicer or 25 nM hAgo2, or with no protein for 30 min at 37 °C. Prior to the addition to the reaction mixtures, the proteins were preincubated either with miRNA or Ex21 (mRNA) for 15 min at 4 °C

Article Snippet: Recombinant human Ago2 (hAgo2) protein was purchased from Active Motif, Giardia intestinalis endoribonuclease Dicer-like recombinant protein (GiDicer) was from MyBiosource, and recombinant human Dicer (hDicer) was produced in our laboratory.

Techniques: Clear Native PAGE, Labeling, Incubation

Journal: Cellular and Molecular Life Sciences

Article Title: The RNA–RNA base pairing potential of human Dicer and Ago2 proteins

doi: 10.1007/s00018-019-03344-6

Figure Lengend Snippet: Genomic features of transcripts for which genomic locations overlapped between shared binding sites of Dicer and Ago2. The values in the chart show the percentage of frequency for distinct genomic features in relation to the total number of shared binding sites (5565). For locations with more than one corresponding transcript, the genomic feature common for the highest number of corresponding transcripts was chosen for calculations

Article Snippet: Recombinant human Ago2 (hAgo2) protein was purchased from Active Motif, Giardia intestinalis endoribonuclease Dicer-like recombinant protein (GiDicer) was from MyBiosource, and recombinant human Dicer (hDicer) was produced in our laboratory.

Techniques: Binding Assay

Journal: Oncotarget

Article Title: Neuropilin-1 is a receptor for extracellular miRNA and AGO2/miRNA complexes and mediates the internalization of miRNAs that modulate cell function

doi: 10.18632/oncotarget.10929

Figure Lengend Snippet: A. miR-20a, pre-complexed or not with 1 nM sNRP1, binds to the plate coated with the Piwi domain of AGO2. Binding is expressed in relative luminescence units (RLU) after the subtraction of non-specific binding (arbitrary units). B. Full- length AGO2 binds to the immobilized NRP1-Fc. AGO2 is pre-mixed or not with the equimolar amount of miR-20a and serially diluted after 1 h incubation. Retention of AGO2 is quantified with anti-AGO2 antibody and visualized with TMB substrate. Binding is expressed as OD 450 -non-specific binding. All binding assays are performed in the presence of Ca 2+ and Mg 2+ . C. ACHN cells internalize miR-331-AGO2 complex conjugated to streptavidin-coated fluorescent beads. Biotin-labeled miR-331 was preincubated with Piwi domain of AGO2 in an equimolar ratio and conjugated to the beads. The internalization of this complex by ACHN cells was observed by confocal microscopy. The beads (green) co-localize with NRP1 (red). Cells are counter-stained for f-actin (cyan). D. Interaction of miRNA with sNRP1, AGO2, or both proteins, facilitates the uptake of miR-331 by ACHN cells. Biotin-labeled miR-331 was preincubated or not with sNRP, AGO2-Piwi, or an equimolar mix of both proteins and conjugated to the beads as in panel C. miR-331 conjugated to the fluorescent streptavidin-coated beads (green) is seen in z-sections cutting through cytoplasm. The cell boundaries are delineated by f-actin staining (grey). E. The uptake of miRNA as shown in panel D is quantified as mean green fluorescence per cell using ImageJ. The data is presented as Mean±SEM; **** P<0.0001 versus all other bars. The data is representative of two independent experiments.

Article Snippet: Human recombinant AGO2 (Piwi domain and a full-length protein) was from Biorbyt and anti-pan-AGO2 antibody was from EMD (Etobicoke, Canada)

Techniques: Binding Assay, Incubation, Labeling, Confocal Microscopy, Staining, Fluorescence