| | |||||||
| Register | Search | Today's Posts | Mark Forums Read |
| Cell Biology and Cell Culture Cell Biology Forum. Cell Culture Forum. Post and ask questions about cell culturing, cell lysis, cell transfection, cell growth, and cell biology. |
| | LinkBack | Thread Tools | Display Modes |
|
#1
| |||
| |||
| Never tried this before, but I'm sure someone out there has done something similar. We could use some advice to prevent us from wasting our $$$. We are using a substrate which contains a fluorogen to detect the presence of our favorite enzyme. As a result of the enzymatic reaction, p-nitroaniline is produced and can be detected via fluorescence. While we have an automated plate-reader that can do fluorescence readings, we need to buy the appropriate filter(s) (for the platereader) to get the proper wavelengths to work with p-nitroaniline. The literature is a bit confusing on this point. Some sources cite an excitation wavelength of 340 nm; others 355 nm; and one even said 405 nm (although this last one might be to detect p-nitroaniline via simple absorbance). If anyone out there is well-versed in fluorescent detection of p-nitroaniline, can you suggest an optimal excitation wavelength (and the cognate emission wavelength)? Thanks, Stan send replies to: [Only registered users see links. ] |
|
#2
| |||
| |||
| John Hines wrote: You could use a fluorescence spectrophotometer, e.g., Hitachi F-2000 or equivalant, to empirically determine the best excitation/emission wavelengths for your samples. Eric |
| Tags |
| detect , fluorescence , paranitroaniline |
| Thread Tools | |
| Display Modes | |
|
|
| | ||||
| Thread | Thread Starter | Forum | Replies | Last Post |
| Single-Molecule Fluorescence Correlation Spectroscopy (FCS) | carlahee | BioPhysics Forum | 1 | 10-18-2010 09:21 AM |
| ayuda para escoger anticuerpo secundario para ELISA | oscardavidcom | Foro Biologia Molecular | 3 | 07-29-2010 10:48 PM |
| Fluorescence And Enzyme & Binding Assays Training Courses | aftabac | Conferences , Symposiums and Meetings | 0 | 12-10-2007 04:38 PM |
| (IVÁN): DECLARADO SANTO PARA DIOS | IVAN VALAREZO | Forum Chemia | 0 | 09-09-2007 01:51 AM |
| detection of beta-nitroaniline via fluorescence? | John Hines | Protein Forum | 0 | 10-22-2003 07:05 PM |