NS340
Milli-Mark® ChromaPan Neuronal Marker-ORC
Milli-Mark ChromaPan Neuronal Marker-ORC is an antibody targeting the ORC protein, validated for use in ICC & IF.
About This Item
Recommended Products
clone
polyclonal
Quality Level
species reactivity
human, mouse, rat
manufacturer/tradename
Milli-Mark®
technique(s)
immunocytochemistry: suitable
immunofluorescence: suitable
shipped in
wet ice
General description
Antibodies to neuronal proteins have become critical tools for identifying neurons and discerning morphological characteristics in culture and complex tissue. 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 has developed a multi-species primary antibody blend that reacts against key somatic, dendritic, and axonal proteins distributed across the neuronal architecture that can then be detected by a single multi-species secondary antibody blend containing discrete fluorophores. As a result, different morphological features (somatic, dendritic, and axonal) are illuminated in different colors: The somato-dendritic cytoarchitecture is detected with DyLight 488 conjugated secondary, and the somato-axonal cytoarchitecture is detected with DyLight 649 conjugated secondary antibody. The primary antibody blend does not contain any rabbit IgGs. The researcher can add a rabbit IgG against their target protein which can be detected with the secondary antibody blend provided by a cyanine 3 conjugated secondary against Rabbit IgG. This pan-neuronal antibody cocktail has been validated in a variety of cell and tissue cultures, giving researchers a convenient and specific qualitative and quantitative tool for studying neuronal morphology.
Specificity
Immunogen
Application
1. Culture neurons as desired. When ready, gently wash with PBS and fix cells with 4% paraformaldehyde in PBS, 5-15 min at room temp (RT).
2. Wash with PBS, 3 x 5min each, at RT and block with 1X blocking buffer for 1 hr at RT.
Note: The blocking buffer provided is supplied as a 5X concentrate. Dilute the concentrate to 1X strength using Milli-Q water. For example, add 20 ml of Milli-Q water to 5 ml of 5X blocking buffer.
3. Incubate cells with ChromaPan Multi-species Primary Antibody blend for 2hr at RT (dilute antibody 1:125 to 1:250 in blocking buffer). Optimal working concentrations must be determined by the end user.
Optional: Rabbit IgG against target protein can also be added to the ChromaPan primary antibody mixture. Optimal working conditions must be determined by the end user. If rabbit IgG is added skip step 4.
4. Wash with PBS, 3 x 5min each, and incubate cells with rabbit IgG against target protein. Optimal working conditions must be determined by the end user.
5. Wash with PBS, 3 x 5min each, and incubate cells with Multi-species Secondary Antibody blend (1:150 in blocking buffer) for 1hr at RT (validated with Chroma Pan to produce minimal background). Optimal working concentrations must be determined by the end user.
Optional: DAPI (1:1000) can also be added to the secondary mixture. If DAPI is added skip step 6.
6. Wash with PBS, 3 x 5min each, and incubate cells with DAPI (1:1000 in blocking buffer) for 5min at RT. Optimal working concentrations must be determined by the end user.
7. Wash with PBS, 3 x 5min each, and cover slip with LIGHT DIAGNOSTICS Mounting Fluid (Catalog No. #5013). View using fluorescent microscope with appropriate filter.
Neuroscience
Neuronal & Glial Markers
Components
2) Multi-species Primary Antibody blend (ORC) (Part No. CS204300): 100uL of a proprietary antibody blend with host specific antibodies that reacts against key somatic, nuclear, dendritic and axonal targets. It does not contain rabbit IgG. Presented in PBS containing 0.05% azide. Store at 2-8°C.
3) Multi-species Secondary Antibody blend (Part No. CS204289): 100 µL of a secondary antibody blend containing host specific IgG antibodies against mouse, rabbit, and chicken, conjugated to DyLight 488, cyanine 3, and DyLight 649 dyes respectively. Each individual set has been species absorbed against H, R, B, Gt, Sh, Gp, Eq and/or M, Rb, Ch to minimize background and cross reactivity. Presented in 0.01M Sodium Phosphate, 0.25M NaCl, pH 7.6 with 15 mg/mL BSA, and 0.05% sodium azide. Store at 2-8°C, away from light.
4) DAPI (Part No. CS202687): Vial contains 10 µL of 0.1mg/mL of DAPI. Store at 2-8°C, away from light.
Quality
Physical form
Storage and Stability
Analysis Note
Cortical neurons
Legal Information
Disclaimer
Signal Word
Warning
Hazard Statements
Precautionary Statements
Hazard Classifications
Aquatic Chronic 3 - Eye Irrit. 2
Storage Class Code
12 - Non Combustible Liquids
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Regulatory Listings
Regulatory Listings are mainly provided for chemical products. Only limited information can be provided here for non-chemical products. No entry means none of the components are listed. It is the user’s obligation to ensure the safe and legal use of the product.
JAN Code
NS340:
Certificates of Analysis (COA)
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