rabbit polyclonal antibody against vamp3  (Thermo Fisher)

Journal: International Journal of Molecular Sciences

Article Title: The Diffusion Model of Intra-Golgi Transport Has Limited Power

doi: 10.3390/ijms24021375

Figure Lengend Snippet: Reagents.

Article Snippet: Rabbit polyclonal antibody against VAMP3 , ThermoFisher Scientific, Milan, Italy , PA5-116218.

Techniques: Recombinant, Transfection, Modification, Plasmid Preparation

Journal: PLoS Pathogens

Article Title: Revisiting Leishmania GP63 host cell targets reveals a limited spectrum of substrates

doi: 10.1371/journal.ppat.1010640

Figure Lengend Snippet: A. Lysates of uninfected BMM (0 h post-infection) or BMM infected with either WT, Δgp63 , or Δgp63 + GP63 L . major metacyclic promastigotes were prepared at the indicated time points in lysis buffer containing 2 mM 1,10-phenanthroline or were lysed directly in 8 M urea. Levels and integrity of Syt XI were assessed by Western blot analysis. β-actin was used a loading control. B. Lysates of uninfected BMM (0 h post-infection) or BMM infected with either WT, Δgp63 , or Δgp63 + GP63 L . major metacyclic promastigotes were prepared at the indicated time points in standard lysis buffer or in lysis buffer containing 10 mM 1,10-phenanthroline. Levels and integrity of VAMP3 and VAMP8 were assessed by Western blot analysis. A low and an high exposure anti-VAMP8 blots are shown. β-actin was used a loading control. Asterisk denotes cleavage fragments. Immunoblots shown are representative of at least four independent experiments. NI, non-infected BMM; HPI, hours post-infection. C. BMM were infected with either WT, Δgp63 , or Δgp63 + GP63 L . major metacyclic promastigotes for 6 h and the distribution of VAMP8 (green) was examined by immunofluorescence confocal microscopy. White arrowheads denote internalized parasites. These results are representative of three independent experiments. DNA is in blue; bar, 10 μM.

Article Snippet: Rabbit polyclonal antibodies against Syntaxin-5, VAMP3, and VAMP8 were obtained from Synaptic Systems; rabbit polyclonal against β-actin, p65RelA, and mTOR from Cell Signaling; rabbit polyclonal anti-c-Jun from Santa Cruz Biotechnology; mouse monoclonal anti-SHP-1 from Abcam; mouse monoclonal antibody anti-NLRP3 from AdipoGen Life Sciences; mouse monoclonal antibodies against Calnexin and Golgin 97 from Invitrogen.

Techniques: Infection, Lysis, Western Blot, Control, Immunofluorescence, Confocal Microscopy

Journal: eLife

Article Title: Synergistic actions of v-SNARE transmembrane domains and membrane-curvature modifying lipids in neurotransmitter release

doi: 10.7554/eLife.55152

Figure Lengend Snippet: ( A ) Exemplary immunofluorescence images of wt and dko cells expressing either GFP, VAMP2 or its TMD mutant variants. Cells were immunolabelled with an affinity purified mouse monoclonal antibody against VAMP2 (clone 69.1, antigen epitope amino acid position 1–14). Note that signals in dko cells expressing VAMP2 or its mutant variants were stronger than in wt+GFP cells and were visualized after adjustment of the camera’s exposure time (e.g. wt+GFP,1.18s; dko+VAMP2, 0.076 s). ( B ) Mean total fluorescence intensity of wt+GFP, dko+GFP and dko cells expressing VAMP2 or the indicated mutants (determined 3.5 hr after transfection). ( C ) Representative structured illumination microscopy images (SIM) for VAMP2 (red) and the other v-SNARE protein VAMP3 (green) in wt and VAMP2-ko chromaffin cells expressing either VAMP2 or the indicated VAMP2-TMD mutant. VAMP2 fluorescence signals of wt cells were illuminated with a threefold higher laser power than in virus-transfected cells. The merged images and the corresponding line scans (dashed lines, right panels) display a clear colocalization between VAMP3 and VAMP2 or its mutant variants. ( D ) Mander’s weighted colocalization of VAMP2 or its TMD mutants to endogenous VAMP3 indicates a similar colocalization coefficient (71.9 ± 2%) for VAMP2 and the TMD mutant variants (VAMP2ko+: VAMP2, 73.8 ± 2%; VAMP2-VAMP1, 78 ± 3%; VAMP2-VAMP8, 76 ± 2%). Numbers of analyzed cells are indicated in the bar. ( E ) The fluorescence intensity of the discrete immunopositive puncta is similar for VAMP2 and the mutant protein and four times higher than for the littermate control (wt) cells. Images were thresholded to values 6 x SD of the background fluorescence to isolate discrete regions of interest for analysis. Data are represented as mean ± SEM. ***p<0.001, one-way analysis of variance between indicated groups followed by Tukey-Kramer post test ( B, D and E ).

Article Snippet: For immunostaining the affinity-purified mouse monoclonal antibody against VAMP2 (clone 69.1, antigen epitope amino acid position 1–14, kindly provided by R. Jahn, MPI for Biophysical Chemistry, Göttingen, Germany) and/or a rabbit polyclonal antibody against VAMP3 (TG-21, Synaptic Systems, Göttingen Germany).

Techniques: Immunofluorescence, Expressing, Mutagenesis, Affinity Purification, Fluorescence, Transfection, Microscopy, Virus, Control

Journal: eLife

Article Title: Synergistic actions of v-SNARE transmembrane domains and membrane-curvature modifying lipids in neurotransmitter release

doi: 10.7554/eLife.55152

Figure Lengend Snippet:

Article Snippet: For immunostaining the affinity-purified mouse monoclonal antibody against VAMP2 (clone 69.1, antigen epitope amino acid position 1–14, kindly provided by R. Jahn, MPI for Biophysical Chemistry, Göttingen, Germany) and/or a rabbit polyclonal antibody against VAMP3 (TG-21, Synaptic Systems, Göttingen Germany).

Techniques: Recombinant, Transfection, Construct, Virus, Derivative Assay, Expressing, Mutagenesis, Software