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.
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Three groups of Poly-L-ornithine coatings are founded to be the most completed supplies:
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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
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Collection of poly l ornithine coating protocols:
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Poly l ornithine coating protocol from CSH
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Prepare 1.5-mg/mL (100X) aliquots of poly-L-ornithine hydrochloride by diluting in H2O. Store at −20°C.
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2. Prepare coated plates:
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To coat 6-well plates for biochemistry:
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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.
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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.
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iii. Store covered in foil at 4°C if the plate is not to be used immediately.
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To coat 24-well plates for morphology assays:
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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.
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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.
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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
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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.
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Aliquot laminin (Cat. #:L2020, Sigma) on ice and keep the aliquots in -80°C until use.
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Coat 6-well plate withpoly-l-ornithine (20μg/mL) 1 mL / well, and incubate at 37°C for 2hours.
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Aspirate poly-l-ornithine and rinse the well twice with 1 mL of DPBS.
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Add 1 mL of lamininsolution (5μg/mL) in the well and incubate at 37°C for 2 hours.
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Aspirate the laminin before plating cells or store plate at 4°C until needed.
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Poly l ornithine coating protocol from Trevigen
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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.
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Slides may be dipped in the solution and air dried before applying sample. Keep sterile.A.
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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.
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Swirl the plate to ensure that it is fully coated.
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Incubate the platefor 1-2 hours at 37°C.
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Remove excess reagent.Rinse the wells three timeswithcell culture medium.
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Plate cells on Poly-L-Ornithine-coated dish
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The List of Poly-L-Ornithine (PLO) + Laminin coating for Neuronal stem cell culture:
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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2. The list of Poly-L-Ornithine + Fibronectin coating for neuron cell culture:
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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3. The List of Poly-L-Ornithine coated German coverslips for cell culture:
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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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