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Poly-L-Ornithine coating

Neuvitro Corporation owns the leading technology to provide professional coating with the broadest coverage of substrates, such as PDL, PLL, PLO, Collagen, Gelatin, Laminin, Fibronectin, PDL+Laminin, PDL + Fibronectin, PLO+Laminin, PLO+Fibronectin, PDL+Laminin+Fibronectin, etc. The majority of coated German coverslips available from Electron Microscopy Science, ThermoFisher Scientific, and VWR is actually manufactured by Neuvitro Corporation.

Poly-L-Ornithine is a synthetic amino acid chain that is positively charged and is widely used as a coating substrate to enhance cell attachment and adhesion to both plastic ware and glass surfaces. Poly-L-Ornithine is often used in combination with laminin for many types of neuronal and neural stem cell cultures. Poly-L-ornithine (PLO) has been used successfully for the attachment and study of neural cell differentiation and outgrowth.

Three groups of Poly-L-ornithine coatings are founded to be the most completed supplies:

1. Poly-L-Ornithine + Laminin coating for neuronal growth and differentiation,

2. Poly-L-ornithine + Fibronectin coating for neuron culture, and

3. Poly-L-Ornithine coated German coverslips for cell culture

Collection of poly l ornithine coating protocols:

Poly l ornithine coating protocol from CSH

  • Prepare 1.5-mg/mL (100X) aliquots of poly-L-ornithine hydrochloride by diluting in H2O. Store at −20°C.

  • 2. Prepare coated plates:

    • To coat 6-well plates for biochemistry:

    • i. Dilute the 100X poly-L-ornithine to 1X in sterile 1X PBS, and pipette enough of the solution to cover the surface of each well. Incubate plates overnight at 37°C.

    • ii. The next day, aspirate the coating solution and wash three times with H2O, making sure the H2O thoroughly fills each well. Air dry in a tissue culture hood until no H2O remains.

    • iii. Store covered in foil at 4°C if the plate is not to be used immediately.

    • To coat 24-well plates for morphology assays:

    • iv. Sterilize a packet of 12-mm coverslips by rinsing for several hours in 70% ethanol and drying overnight under the UV light of the tissue culture hood.

    • v. Place one coverslip in each well of the 24-well plate, add 0.5 mL of 1X poly-L-ornithine coating solution (diluted as in Step 2.i) per well, and follow the incubation/rinsing procedure described in Step 2.ii, using at least 1 mL of H2O per well.

Poly l ornithine (PLO) coating protocol from Sigma

Optimal coating concentrations must be determined by end user. Typical coating concentrations range from 10ug/ml to 100ug/ml based on cell type and application.

1. Thaw Poly-L-Ornithine solution at room temperature.
2. Dilute Poly-L-Ornithine solution to desired concentration in sterile water.
3. Fully coat the cell culture surface with diluted Poly-L-Ornithine solution. Use 5 mL volume for 6-cm plates and 10 mL volume for 10-cm plates and T75 flasks.
4. Allow cell culture vessel to sit at room temperature overnight.
5. Aspirate the Poly-L-Ornithine solution the following day and rinse vessel with sterile water followed by coating with desired ECM protein.

 

Poly l ornithine coating protocol from 101bio

  1. Dilute poly-l-ornithine (Cat.#: P3655, Sigma) on ice using sterile tissue culture-grade water(20μg/mL). Aliquot the diluted poly-l-ornithin and keep the aliquots in -80°C until use.

  2. Aliquot laminin (Cat. #:L2020, Sigma) on ice and keep the aliquots in -80°C until use.

  3. Coat 6-well plate withpoly-l-ornithine (20μg/mL) 1 mL / well, and incubate at 37°C for 2hours.

  4. Aspirate poly-l-ornithine and rinse the well twice with 1 mL of DPBS.

  5. Add 1 mL of lamininsolution (5μg/mL) in the well and incubate at 37°C for 2 hours.

  6. Aspirate the laminin before plating cells or store plate at 4°C until needed.

Poly l ornithine coating protocol from Trevigen

  1. The recommended working concentration is 0.1 mg/mL (as provided),but optimal conditions for attachment must be determined for each cell line and application.

  2. Slides may be dipped in the solution and air dried before applying sample. Keep sterile.A.

  3. The following table is a guide for the suggested volumes required:Plate TypeVolume Poly-L-Ornithine/Well10 cm dish (or 75 cm2flask)10 mL6 cm dish(or 25 cm2flask)3-5mL6 wells (or 35 mm dish) 1 mL12 well500 μLB.Pipette the appropriate amount of Poly-L-Ornithinesolution in each well.

  4. Swirl the plate to ensure that it is fully coated.

  5. Incubate the platefor 1-2 hours at 37°C.

  6. Remove excess reagent.Rinse the wells three timeswithcell culture medium.

  7. Plate cells on Poly-L-Ornithine-coated dish

The List of Poly-L-Ornithine (PLO) + Laminin coating for Neuronal stem cell culture:

Cat. #: GG-12-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 12mm diameter #1 thickness 60 pieces sterile. Shelf life 1/2 year.

Cat. #: GG-12-1.5-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 12mm diameter #1.5 thickness 60 pieces sterile ready to use in cell culture. Shelf life 1/2 year.

Cat. #: GG-14-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 14mm diameter #1 thickness 60 pieces sterile ready to use in cell culture. Shelf life 1/2 year.

Cat. #: GG-15-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 15mm diameter #1 thickness 60 pieces sterile. Shelf life 1/2 year.

Cat. #: GG-18-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 18mm diameter #1 thickness 60 pieces sterile, ready to use. Shelf life 1/2 year.

Cat. #: GG-18-1.5-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 18mm diameter #1.5 thickness 60 pieces sterile. Shelf life 1/2 year.

Cat. #: GG-20-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 20mm diameter #1 thickness 60 pieces sterile. Shelf life 1/2 year.

Cat. #: GG-22-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 22mm diameter #1 thickness 60 pieces sterile. Shelf life 1/2 year.

Cat. #: GG-22-1.5-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 22mm diameter #1.5 thickness 60 pieces sterile. Shelf life 1/2 year.

Cat. #: GG-24-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 24mm diameter #1 thickness 60 pieces sterile. Shelf life 1/2 year.

Cat. #: GG-25-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 25mm diameter #1 thickness 60 pieces sterile. Shelf life 1/2 year.

Cat. #: GG-25-1.5-PLO+Laminin

Item description: Laminin coating over Poly-L-ornithine layer on German coverglasses to promote neuronal stem cell growth and differentiation. 25mm diameter #1.5 thickness 60 pieces sterile, ready to use. Shelf life 1/2 year.

2. The list of Poly-L-Ornithine + Fibronectin coating for neuron cell culture:

Cat.#: GG-12-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 12mm diameter, #1 thickness, 60 pieces, sterile, ready to use in multi-well culture plate.

Cat.#: GG-12-1.5-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 12mm diameter, #1.5 thickness, 60 pieces, sterile, ready to use in multi-well culture plate.

Cat.#: GG-14-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 14mm diameter, #1 thickness, 60 pieces, sterile, ready to use in multi-well culture plates.

Cat.#: GG-15-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 15mm diameter, #1 thickness, 60 pieces, sterile, ready to use in multi-well culture plates.

Cat.#: GG-18-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 18mm diameter, #1 thickness, 60 pieces, sterile, ready to use in multi-well culture plate.

Cat.#: GG-18-1.5-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 18mm diameter, #1.5 thickness, 60 pieces, sterile, ready to use in multi-well culture plate.

Cat.#: GG-20-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 20mm diameter, #1 thickness, 60 pieces, sterile, ready to use in multi-well culture plate.

Cat.#: GG-22-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 22mm diameter, #1 thickness, 60 pieces, sterile, ready to use in multi-well culture plate.

Cat.#: GG-22-1.5-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 22mm diameter, #1.5 thickness, 60 pieces, sterile, ready to use in multi-well culture plate.

Cat.#: GG-24-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 24mm diameter, #1 thickness, 60 pieces, sterile, ready to use in multi-well culture plate.

Cat.#: GG-25-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 25mm diameter, #1 thickness, 60 pieces, sterile, ready to use in multi-well culture plate.

Cat.#: GG-25-1.5-PLO+Fibronectin

Item description: Fibronectin coating on Poly-L-ornithine coated German coverslip for neuron culture and cell culture, 25mm diameter, #1.5 thickness, 60 pieces, sterile, ready to use in multi-well culture plate.

3. The List of Poly-L-Ornithine coated German coverslips for cell culture:

Cat. #: GG-12-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 12mm diameter #1 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-12-1.5-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 12mm diameter #1.5 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-14-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 14mm diameter #1 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-15-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 15mm diameter #1 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-18-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 18mm diameter #1 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-18-1.5-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 18mm diameter #1.5 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-20-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 20mm diameter #1 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-22-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 22mm diameter #1 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-22-1.5-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 22mm diameter #1.5 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-24-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 24mm diameter #1 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-25-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 25mm diameter #1 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

Cat. #: GG-25-1.5-PLO

Item description: Poly-L-Ornithine coated German coverglasses for cell culture, 25mm diameter #1.5 thickness, 90 pieces, sterile, ready to use in well plates, shelf life 1/2 year.

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