Neuro-Chrom ChromaPan Neuronal MarkerORC

Neuro-Chrom ChromaPan Neuronal MarkerORC

Stock Code: 3587269
Manufacturer Part No: NS340
Order Now for 7 day delivery
Click Add to Quote for price and delivery
Quantity: - +

Analysis Note


ControlCortical neurons


Application


Research Sub CategoryNeuronal & Glial Markers


Research CategoryNeuroscience


Immunocytochemistry Protocol
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.



Components


1) 5X Blocking Buffer (Part No. CS204291): 5 ml of an optimized blocking buffer is provided and contains PBS, BSA, serum, trition X-100, and 0.05% azide. Store at 2-8°C.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.


Disclaimer


Quality Level100
ManufacturerSIGMA-ALDRICH
Technique(s)immunocytochemistry: suitable, immunofluorescence: suitable

There are no downloads for this product.