24-well tissue culture plates were coated with 1X Poly-L-Ornithine (Corning Inc., Lowell, MA, 25–055-CV). 2.5×105 Shh-LIGHT2 cells17 (link) were plated per well in 500µl media containing DMEM with 10%FBS and 1% P/S. 24 hours later, cells were changed to DMEM with 0.5% FBS and transduced with red fluorescent protein (RFP)-tagged control (RFP-CTRL) or yellow fluorescent protein (YFP)-tagged WIP1 (YFP-WIP1) lentivirus. 24 hours later, cells were stimulated with vehicle or Shh (3µg/mL) for 24 hours. Luciferase activity was assayed using the Dual Luciferase Reporter Assay System (Promega, Madison, WI, # E1910), according to manufacturer’s instructions.
Poly l ornithine
Manufactured by Corning
Sourced in United States
Poly-L-Ornithine is a synthetic polymer of the amino acid L-ornithine. It is commonly used as a cell culture coating to promote cell attachment and growth in various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Dual luciferase reporter assay system, by Promega (2 mentions)
Axiovert 200m, by Zeiss (2 mentions)
Axiovision 4, by Zeiss (1 mentions)
Axiocam hrm, by Zeiss (1 mentions)
24 well plate, by Corning (1 mentions)
Tempcontrol 37 2 digital, by Pecon (1 mentions)
Cti controller 3700 digital, by Pecon (1 mentions)
3 protocols using poly l ornithine
1
Shh-Responsive Luciferase Assay
2
Shh-Responsive Luciferase Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
24-well tissue culture plates were coated with 1X Poly-L-Ornithine (Corning Inc., Lowell, MA, 25–055-CV). 2.5×105 Shh-LIGHT2 cells17 (link) were plated per well in 500µl media containing DMEM with 10%FBS and 1% P/S. 24 hours later, cells were changed to DMEM with 0.5% FBS and transduced with red fluorescent protein (RFP)-tagged control (RFP-CTRL) or yellow fluorescent protein (YFP)-tagged WIP1 (YFP-WIP1) lentivirus. 24 hours later, cells were stimulated with vehicle or Shh (3µg/mL) for 24 hours. Luciferase activity was assayed using the Dual Luciferase Reporter Assay System (Promega, Madison, WI, # E1910), according to manufacturer’s instructions.
3
Time-lapse Microscopy of NSPC Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For time-lapse video microscopy, the neurospheres were dissociated as described in section “2.4 Differentiation and proliferation assay under differentiation conditions.” The NSPCs were plated in a density of 30000 cells per well on poly-L-ornithine (15 μg/ml) coated 24 well plates (Corning, Corning, NY, USA, Cat# 353226) in neurosphere medium. After 2 h, the cells adhered and the medium was supplemented with EGF, FGF2 and heparin in the same concentrations as in the neurosphere culture to ensure proliferation conditions. For the analysis, the produced FnIII Fc recombinants were presented to the cells as coated substrates or as soluble medium additives, both with a concentration of 25 μg/ml. A total of 2 h after seeding the plate was placed in the video microscope system of an Axiovert 200 M provided with an AxioCam HRm and AxioVision-4.8.1 software (Carl Zeiss, Oberkochen, Germany). Additionally, two regulating elements, namely the “Tempcontrol 37-2 digital” and the “CTI-Controller 3700 digital” (PeCon GmbH, Erbach, Germany) ensured stable temperature and pH conditions. Within the system the cells were cultured at 37°C and 5% CO2. For analysis, five spots per domain combination were chosen and a picture was taken every 6 min for 96 h.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!