Abcam
Rabbit anti-Calpain-1
ab39170
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Stress in vivo, and CRH application in vitro, cause dendritic spine loss in hippocampal pyramidal neurons. This loss of spines, and subsequent synapses, is likely the underlying mechanism through which severe stress disrupts learning and memory. CRH, in combination with neuronal activity and NMDA receptor activation, recruits the actin destabilizing protein, calpain, to break down the actin cytoskeleton resulting in spine loss. Identifying the downstream signaling from CRH receptors could reveal potential therapeutic targets for diseases such as post-traumatic stress disorder (PTSD), anxiety, and depression.Calpain is a calcium-activated protease that is important for neuronal processes such as long-term potentiation (LTP), spine actin remodeling, and receptor cleavage. Calpain has recently been identified as an important protease involved in stress- and CRH-induced dendritic remodeling.
Immunofluorescence
Dissociated hippocampal neurons
1:1000
3% BSA + 0.1% Triton-X100 in PBS
1:400
None
Fluorescence
I performed a serial dilution of the antibody and tried 1:250, 1:500, 1:1000, and 1:2000. For dissociated hippocampal neurons grown in culture till day in vitro 17 (DIV17) 1:1000 produced a punctate immunocytochemistry. This is consistent with the literature localizing calpain in the hippocampus and to dendritic spines.
Washing with PBS + 0.1% Triton after the secondary antibody incubation seemed to improve background
Antibody looks specific and produces the punctate immunocytochemistry consistent with proteins localized to dendritic spines
I would recommend this antibody for neurons and other cell types.