Analysis Note

ControlCortical neurons

Application

Research Sub CategoryNeuronal & Glial Markers

Research CategoryNeuroscience

Immunocytochemistry:

1. Culture neurons as desired. When ready, gently wash with PBS and fix cells with 4% paraformaldehyde in PBS, 30min at room temp (RT).
2. Wash with PBS, 3 x 5min each, at RT and block with blocking buffer (1% BSA, 5% Serum, 0.2% Triton X in PBS) for 1 hr at RT.
3. Incubate cells with Milli-Mark FluoroPan Neuronal Marker (Mouse IgG conjugated with Alexa 488) for 2hr at RT (dilute antibody in blocking buffer).
4. Wash with PBS, 3 x 5min each, and cover slip with LIGHT DIAGNOSTICS Mounting Fluid (Catalog No. #5013). View using fluorescent microscope with FITC filter.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Antibodies to neuronal proteins have become critical tools for identifying neurons and discerning morphological characteristics in culture and complex tissue. While the labeling from classic histological techniques, such as Golgi staining, and modern molecular approaches, such as GFP constructs, yield excellent cytoarchitectural detail, these approaches are technically challenging and impractical for many neuroscience research needs. Neuron-specific antibodies are convenient precision tools useful in revealing cytoarchitecture, but are limited to the protein target distribution within the neuron, which may differ greatly from nucleus to soma to dendrite and axon. To achieve as complete a morphological staining as possible across all parts of neurons, Millipore previously developed a monoclonal antibody blend (Catalog No. MAB2300) that reacts against key somatic, nuclear, dendritic, spine, and axonal proteins distributed across the pan-neuronal architecture. Now Millipore has formulated MAB2300X, an Alexa 488 conjugated version of the pan-neuronal antibody blend MAB2300 to facilitate immunofluorescence studies. Each antibody within the blend is conjugated to Alexa 488 therefore enabling detection of an entire neuron with only one microscopic emission channel. MAB2300X has been validated in diverse fixations, cell culture, and immunohistochemistry protocols, giving researchers a convenient and specific qualitative and quantitative tool for studying neuronal morphology. High morphological resolution has been achieved using MAB2300x including distral dentrites and spines.

Immunogen

Epitope: Whole Neuron Marker

Legal Information

ALEXA FLUOR is a registered trademark of Life Technologies

MILLI-MARK is a registered trademark of Merck KGaA, Darmstadt, Germany

Physical form

Blended monoclonal antibody cocktail in PBS with 0.05% NaN3

Quality

Routinely tested on cortical neurons in Immunocytochemistry 1:150 - 1:100.

Specificity

MAB2300X is specific to axons (neurites), dendrites, spines, nucleus, and the cell body of neurons.

Reactivity with other species has not been determined.

Storage and Stability

Maintain at 2-8°C for up to 6 months from date of receipt.