rabbit anti-calbindin d28 antibody  (Synaptic Systems)

Journal: bioRxiv

Article Title: Calbindin-containing CA1 pyramidal cells support cognitive flexibility in spatial task in mice

doi: 10.64898/2025.11.30.691413

Figure Lengend Snippet: This figure highlights the progressive optimization of CALB1 staining, with the most effective staining observed in panel (C and E). All representative images show stainings of CALB1 in red and DAPI in blue. (A) Original Protocol: Representative images of stainings using the original protocol . Blocking solution without glycine (PBS, 0.1% Triton X-100, 5% NGS). Primary antibody solution contains: PBS, 0.1% Triton X-100, 2.5% NGS, 1:1000 rabbit anti-Calbindin D28k antibody (CB-38a; Swant, Switzerland; 1:2000). Secondary antibody solution contains: PBS, 0.1% 0.1% Triton X-100, 0.5% Tween20, Alexa Fluor anti-rabbit antibody (1:1000). Washing buffer contains: 0.5% Tween20. Shown are an overview image of the mouse brain with focus on the dorsal hippocampus (2.5x), the cortex (40x), and the CA1 pyramidal cell layer (20x). (B) 1hr incubation with 50mM Glycine in blocking buffer: Representative images of stainings using 50mM Glycine in blocking buffer, which is kept on for 1hr. The primary antibody solution also contains 10 mM glycine. Images show overview of dorsal hippocampus (10x), cortex (20x), and CA1 pyramidal cell layer (20x). (C) Overnight incubation with 50mM Glycine in blocking buffer: Blocking performed overnight with 50 mM glycine; the primary antibody solution also contains 10 mM glycine. Shown are an overview of the dorsal hippocampus (10x) and the CA1 pyramidal cell layer (40x). (D) 1hr incubation with 50mM Glycine in blocking buffer and antigen-signal enhancement (ASE) protocol of primary antibody . Representative images of stainings using 50 mM glycine in blocking buffer for 1hr; primary antibody solution supplemented with ASE (10 mM glycine, 0.1% H 2 O 2 , 0.05% Tween20, 0.1% Triton X-100). Shown are an overview of the dorsal hippocampus (20x) and the CA1 pyramidal cell layer (40x). (E) Overnight incubation with 50mM Glycine in blocking buffer and ASE protocol: Representative images of stainings using 50 mM glycine in blocking buffer overnight; primary antibody solution according to ASE protocol. Shown are an overview image of the mouse brain with focus on the dorsal hippocampus (2.5x), and the CA1 pyramidal cell layer at 10x and 20x.

Article Snippet: To address this, we focused on identifying visible changes in staining intensity and distribution using the antibody rabbit anti-Calbindin D28 K (CB-38a; Swant, Switzerland; 1:2000).

Techniques: Staining, Blocking Assay, Incubation

Journal: Neurobiology of Stress

Article Title: Persistent pain intensifies recall of consolidated fear memories

doi: 10.1016/j.ynstr.2019.100163

Figure Lengend Snippet: Ongoing pain induced morphological changes in the BLA interneuronal population. Panels A and B show calbindin immunoreactivity (Calb-ir, green) and panels D and E show parvalbumin immunoreactivity (Parv-ir, red) expression in sham mice ( A and D ) and in cuffed mice ( B and E ). Panels G and H show the combined expression of the two markers in sham ( G ) and cuffed ( H ) mice, where the Calb/Parv-ir co-localization appears as yellow. The number of Calb-ir cells did not differ between the groups ( C) but the number of Parv immunopositive neurons increased in the cuffed mice ( F ) and most of these Parv-ir neurons were colocalized with Calb-ir, or the yellow labeled neurons in the G and H inserts; The graph in panel I compares the number of Calb/Parv-ir neurons of sham and cuffed mice. The cell counts were evaluated by T-test, * p < 0.05 and *** p < 0.001, bars represent mean ± SEM, n = 12 to 14 per group. BLA - basolateral amygdala, CeA - central amygdala, LA - lateral amygdala. Scale bar = 200 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: A similar immunostaining protocol was used for visualization of calbindin and parvalbumin in the BLA using rabbit anti-calbindin D28 antibody (Synaptic Systems, Goettingen, Germany) diluted 1:10 K and monoclonal mouse antibody against parvalbumin (Sawnt, Marly, Switzerland) diluted 1:5 K. The next step was incubation with secondary anti-rabbit Alexa 488 and anti-mouse Alexa 594 antibodies (Jackson ImmunoResearch Inc. West Grove, PA) for 4 h at room temperature.

Techniques: Expressing, Labeling