immunolabeling chicken polyclonal  (Novus Biologicals)

Journal: bioRxiv

Article Title: Endogenous tagging of Munc13-1 with a SNAP tag as a tool for monitoring synapse nanoarchitecture

doi: 10.1101/2024.10.08.617143

Figure Lengend Snippet: Example images of cultured hippocampal neurons from WT (A) and Unc13a SNAP/SNAP mouse brains (B) , immunolabeled with antibodies against Munc13-1, the active zone marker Bassoon, and MAP2 (Scale bar: 20 µm). (C) Quantification of the fluorescence signal arising from antibody labeling of Munc13-1 and (D) of the fluorescence signal arising from antibody labeling of Bassoon in neurons from WT and Unc13a SNAP/SNAP mice from images as in A and B. (E) The colocalization of Munc13-1 and Bassoon signals evaluated by calculating the Pearson’s correlation coefficient.

Article Snippet: Subsequently, the cells were extensively washed up to eight times in blocking solution and the following antibodies were applied in blocking buffer for immunolabeling: Chicken polyclonal MAP2 (Novus Biologicals NB300-213, diluted 1:1000), Rabbit polyclonal Munc13-1 (Synaptic Systems AB_126103, diluted 1:500), and Guinea Pig polyclonal Bassoon (Synaptic Systems AB_141004, diluted 1:500).

Techniques: Cell Culture, Immunolabeling, Marker, Fluorescence, Antibody Labeling

Journal: bioRxiv

Article Title: Endogenous tagging of Munc13-1 with a SNAP tag as a tool for monitoring synapse nanoarchitecture

doi: 10.1101/2024.10.08.617143

Figure Lengend Snippet: (A) Chemical structures of silicone rhodamine variants for SNAP tag labeling, including SBG-SiR-d12 developed here. (B-C) Example images of fixed cultured hippocampal neurons from WT (B) and Unc13a SNAP/SNAP (C) mouse brains immunolabeled with the SNAP tag compounds BG-JF 646 , SBG-JF 646 , BG-SiR-d12, SBG-SiR-d12, as well as with antibodies against Munc13-1, Bassoon, and MAP2 (Scale bar: 20 µm). (D) Quantification of the fluorescence signal arising from Munc13-1-SNAP labeling by BG-JF 646 , SBG-JF 646 , BG-SiR-d12, SBG-SiR-d12, in neurons from WT and Unc13a SNAP/SNAP mice. (E) Colocalization of signals arising from simultaneous Munc13-1 antibody labeling and SNAP tag labeling using the Person’s correlation coefficient.

Article Snippet: Subsequently, the cells were extensively washed up to eight times in blocking solution and the following antibodies were applied in blocking buffer for immunolabeling: Chicken polyclonal MAP2 (Novus Biologicals NB300-213, diluted 1:1000), Rabbit polyclonal Munc13-1 (Synaptic Systems AB_126103, diluted 1:500), and Guinea Pig polyclonal Bassoon (Synaptic Systems AB_141004, diluted 1:500).

Techniques: Labeling, Cell Culture, Immunolabeling, Fluorescence, Antibody Labeling