anti stx16  (Novus Biologicals)

Journal: Virology

Article Title: Integrated proteomics and transcriptomics analyses identify novel cell surface markers of HIV latency

doi: 10.1016/j.virol.2022.06.003

Figure Lengend Snippet: Antibodies used to test enrichment of latently infected cells from mixed cell populations.

Article Snippet: Antibody Isotype Secondary a Conjugate Manufacturer (cat. #) CEACAM1 Mouse IgG2b N/A PE R&D Systems (FAB2244P) PLXNB1 Mouse IgG2a N/A APC R&D Systems (FAB53291A) STX16 Rabbit IgG Goat anti-Rabbi APC Thermofisher Scientific (PA5-48340) LIMK1 Mouse IgG2b Goat anti-Mouse PE Novus Biologicals (NBP2-00748) LAX1 Rabbit IgG Goat anti-Rabbit APC Abcam (ab133759) ANXA2 Rabbit IgG N/A PE Abcam (ab210729) LGALS1 Mouse IgG1 Goat anti-Mouse PE Thermofisher Scientific (43–7400) EPHA2 Mouse IgG2a N/A APC R&D Systems (FAB3035) CD80 Mouse IgG1 N/A FITC R&D Systems (FAB140F-025) CCL5 Mouse IgG1 N/A APC R&D Systems (IC278A) MX1 Rabbit IgG Goat anti-Rabbit APC Abcam (ab207414) KRAS Mouse IgG2b Goat anti-Mouse PE Novus Biologicals (NBP2-59413) Open in a separate window CEACAM1 , CEA cell adhesion molecule 1; PLXNB2 , plexin B2; STX16 , syntaxin 16; LIMK1 , LIM domain kinase 1; LAX1 , lymphocyte transmembrane adaptor 1; ANXA2 , annexin A2; LGALS1 , galectin 1; EPHA2 , EPH receptor A2; CD80 , CD80 molecule; CCL5 , C─C motif chemokine ligand 5; MX1 , MX dynamin like GTPase 1; KRAS , KRAS proto-oncogene, GTPase; PE, phycoerythrin; APC, allophycocyanin; FITC, fluorescein isothiocyanate; cat. #, catalog number; N/A, not applicable. a Secondary antibody is not applicable for primary antibodies conjugated to fluorophores.

Techniques: Infection

Journal: The Journal of Cell Biology

Article Title: MT1-MMP recruits the ER-Golgi SNARE Bet1 for efficient MT1-MMP transport to the plasma membrane

doi: 10.1083/jcb.201808149

Figure Lengend Snippet: Bet1 forms novel SNARE complexes with endosomal SNAREs in MDA-MB-231 cells. (A) 3xFLAG-Bet1 interacts with STX4, Vti1b, and VAMP4 in addition to STX5. MDA-MB-231 cells stably expressing 3xFLAG-Bet1 were lysed, and the lysate was subjected to immunoprecipitation using FLAG-beads, and then the precipitate was subjected to immunoblotting for endogenous SNARE proteins. In the following blots, the brightness and contrast were adjusted so that their input bands became comparable to those of other SNAREs: STX3, STX7, STX16, STX17, STX18, VAMP3, VAMP7, and Vti1b. (B) An endogenous complex comprised of STX4, Bet1, Vti1b, and VAMP4 exists in MDA-MB-231 cells, but not in HeLa cells. Lysates of MDA-MB-231 and HeLa cells were subjected to immunoprecipitation of STX4, and then the co-precipitation of Bet1, Vti1b, and VAMP4 was analyzed. In the following blots, the brightness and contrast were adjusted so that their input bands became comparable to those of other SNAREs: Bet1 and VAMP4 in MDA-MB-231 cells and Bet1, VAMP4, and VAMP8 in HeLa cells. (C and D) Bet1-interacting SNAREs are colocalized with Bet1-GFP in endomembrane compartments. MDA-MB-231 cells stably expressing Bet1-GFP were transfected with FLAG-SNAREs and then subjected to immunofluorescence and confocal microscopy (C). Their colocalization was assessed by using the Manders’ overlap coefficient (D). (E–G) Depletion of STX4 and Vti1b reduces the complex of Bet1 with Vti1b and STX4, respectively. MDA-MB-231 cells stably expressing 3xFLAG-Bet1 were transfected with STX4 and Vti1b siRNAs, and then immunoprecipitation and immunoblotting (E) were performed as in A. Co-precipitated endogenous STX4 (F) and Vti1b (G) were quantified. Scale bar: 10 μm in a regular image; 2 μm in an inset. **, P < 0.01; vs. STX4 in D; vs. FLAG (Bet1), mock in F and G.

Article Snippet: The commercial rabbit polyclonal antibodies used were anti-FLAG (Sigma-Aldrich), anti–MT1-MMP (EMD Millipore), anti-STX16 (Synaptic Systems), anti-GFP (Thermo Fisher Scientific), and anti-GPP130 (Covance).

Techniques: Stable Transfection, Expressing, Immunoprecipitation, Western Blot, Transfection, Immunofluorescence, Confocal Microscopy