Journal: Nature immunology

Article Title: The receptor DNGR-1 signals for phagosomal rupture to promote cross-presentation of dead cell-associated antigens

doi: 10.1038/s41590-020-00824-x

Figure Lengend Snippet: Abbreviations: FC (flow cytometry), IF (immunofluorescence), CI (cell isolation)

Article Snippet: Dectin-1 , monoclonal rat (2A11) , 1mg/ml , IF 1:100 FC 1:100 , GeneTex GTX41467.

Techniques: Flow Cytometry, Immunofluorescence, Isolation, Labeling, Generated, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Human AKTIP interacts with ESCRT proteins and functions at the midbody in cytokinesis

doi: 10.1101/2020.01.19.911891

Figure Lengend Snippet: (A) AKTIP localization at the midbody by immunofluorescence in HeLa cells using anti-AKTIP antibodies WH0064400M2 clone 2A11 (top panel) and HPA041794 (lower panel). (B) Detection of exogenous AKTIP-FLAG by immunofluorescence using anti-FLAG antibody. (C-E) Immunofluorescence with anti-AKTIP (WH0064400M2 clone 2A11) (C) and qPCR (D) showing that AKTIP reduction causes a drop to 47% AKTIP positively staining midbodies as opposed to 91.7% of control cells (E). Results shown are the mean value of two replicates ± SEM ***p < 0.001; Student’s t-test; 60 midbodies per condition were analyzed. Scale bar 5μm.

Article Snippet: To validate the specificity of the localization of AKTIP to the midbody, we compared the immunostaining using the monoclonal anti-AKTIP antibody (2A11 WH006400M2 Abnova, and upper panel) to that obtained with a polyclonal antibody (HPA041794 Sigma; Fig S1A lower panel).

Techniques: Immunofluorescence, Staining, Control

Journal: Chembiochem : a European journal of chemical biology

Article Title: Rapid Development of a Potent Photo-Triggered Inhibitor of the Serine Hydrolase RBBP9

doi: 10.1002/cbic.201200445

Figure Lengend Snippet: Screening Strategy of ABPP Probe facilitated Peptoid Library Screening against Serine Hydrolases. RBBP9 was allowed to react with FP-biotin to yield the active site biotinylated protein. The FP-biotinylated RBBP9 was incubated with OBOC peptoid library. The hit compounds that bound to RBBP9 can be pull out with Streptavidin-coated magnetic beads by Streptavidin-biotin interaction.

Article Snippet: The membrane was blotted with primary anti-RBBP9 monoclonal antibody (Novus Biologicals) followed by secondary conjugated antibody (goat anti-mouse IgG horseradish peroxidase, Bio-Rad).

Techniques: Library Screening, Incubation, Magnetic Beads

Journal: Chembiochem : a European journal of chemical biology

Article Title: Rapid Development of a Potent Photo-Triggered Inhibitor of the Serine Hydrolase RBBP9

doi: 10.1002/cbic.201200445

Figure Lengend Snippet: Activity based labeling of RBBP9 with biotinylated fluorophosphonate probe. A) General mechanism for covalent modification of serine hydrolases active site by fluorophosphonate-biotin. B) Whole protein mass spectra for unmodified and FP-biotin modified RBBP9. C) Gel analysis for the recognition of FP-biotinylated RBBP9 by Streptavidin.

Article Snippet: The membrane was blotted with primary anti-RBBP9 monoclonal antibody (Novus Biologicals) followed by secondary conjugated antibody (goat anti-mouse IgG horseradish peroxidase, Bio-Rad).

Techniques: Activity Assay, Labeling, Modification

Journal: Chembiochem : a European journal of chemical biology

Article Title: Rapid Development of a Potent Photo-Triggered Inhibitor of the Serine Hydrolase RBBP9

doi: 10.1002/cbic.201200445

Figure Lengend Snippet: Mass spectra of on-bead tryptic digestion from Streptavidin-coated magnetic beads after incubation with RBBP9. A) Streptavidin-coated magnetic beads were incubated with unmodified RBBP9. B) Streptavidin-coated magnetic beads were incubated with FP-biotinylated RBBP9. C) Mass spectra of a standard in-solution RBBP9 digestion.

Article Snippet: The membrane was blotted with primary anti-RBBP9 monoclonal antibody (Novus Biologicals) followed by secondary conjugated antibody (goat anti-mouse IgG horseradish peroxidase, Bio-Rad).

Techniques: Magnetic Beads, Incubation

Journal: Chembiochem : a European journal of chemical biology

Article Title: Rapid Development of a Potent Photo-Triggered Inhibitor of the Serine Hydrolase RBBP9

doi: 10.1002/cbic.201200445

Figure Lengend Snippet: Comparison of inhibition potency of the chromophore-hit conjugates in the absence or presence of photo-activation. A) ABPP gel assay showing only light activated Ru(II)-6 blocks RBBP9 labeling by FP-Rh in a concentration dependent manner. B) ABPP gel assay showing only light activated Eosin-1 blocks RBBP9 labeling by FP-Rh in a concentration dependent manner.

Article Snippet: The membrane was blotted with primary anti-RBBP9 monoclonal antibody (Novus Biologicals) followed by secondary conjugated antibody (goat anti-mouse IgG horseradish peroxidase, Bio-Rad).

Techniques: Comparison, Inhibition, Activation Assay, Labeling, Concentration Assay

Journal: Chembiochem : a European journal of chemical biology

Article Title: Rapid Development of a Potent Photo-Triggered Inhibitor of the Serine Hydrolase RBBP9

doi: 10.1002/cbic.201200445

Figure Lengend Snippet: Photo-inactivated RBBP9 by Ru(II)-6 does not react with ABPP probe. A) The photo-inactivated RBBP9 does not response to Coomassie blue staining. Silver staining and Streptavidin-HRP blotting suggest that photo-damaged RBBP9 does not react with FP-biotin. B) Mass Spectra of RBBP9 labeling with FP-Biotin after incubation with Ru(II)-6 in the absence of presence of light activation. C) No endogenous level of RBBP9 was detected by western-blotting with anti-RBBP9 antibody. For the recombinant RBBP9 and RBBP9 doped in cell proteome complex, a ladder of higher molecular weight bands observed with increased Ru(II)-6 upon light activation.

Article Snippet: The membrane was blotted with primary anti-RBBP9 monoclonal antibody (Novus Biologicals) followed by secondary conjugated antibody (goat anti-mouse IgG horseradish peroxidase, Bio-Rad).

Techniques: Staining, Silver Staining, Labeling, Incubation, Activation Assay, Western Blot, Recombinant, Molecular Weight

Journal: Chembiochem : a European journal of chemical biology

Article Title: Rapid Development of a Potent Photo-Triggered Inhibitor of the Serine Hydrolase RBBP9

doi: 10.1002/cbic.201200445

Figure Lengend Snippet: Selective inhibition of RBBP9 in cellular proteome. Evaluation of A) Ru(II)-6, B) Eosin-1 by competitive ABPP in soluble proteome of HeLa cells (1 mg/mL). Recombinant human RBBP9 (400 nM) was doped into this proteome for comparison.

Article Snippet: The membrane was blotted with primary anti-RBBP9 monoclonal antibody (Novus Biologicals) followed by secondary conjugated antibody (goat anti-mouse IgG horseradish peroxidase, Bio-Rad).

Techniques: Inhibition, Recombinant, Comparison