User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/13

From OpenWetWare

< User:Karmella Haynes | Notebook | PcTF Genomics | 2013 | 06(Difference between revisions)
Jump to: navigation, search
(Autocreate 2013/06/13 Entry for User:Karmella_Haynes/Notebook/PcTF_Genomics)
Current revision (19:46, 17 June 2013) (view source)
(mm/dd/yy)
 
Line 8: Line 8:
-
==mm/dd/yy==
+
==06/13/13==
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
-
* Line item 1
+
* Gal4-EED/luc cells IFC - secondary stain & imaging
 +
 
----
----
-
'''Line item 1'''<br>
+
'''Gal4-EED/luc cells IFC'''<br>
-
> Samples
+
* Samples: first row of wells in the 24-well plate.
 +
* Keep all others under PBS
 +
 
 +
# 0 μg/mL dox
 +
# 0.1
 +
# 0.2
 +
# 0.5
 +
# 1.0
 +
# 2.0
 +
 
 +
* Follow standard protocol; wash 3x with 1xPBS
 +
* Use 1:1000 dilution of anti-rabbit Alexa488 and 1:1000 Hoescht (in 15 μL Gentex horse serum blocking solution per well)
 +
* Incubate under parafilm "covers" for one hour at room temperature in light-tight box with wet paper towels
 +
* Use GFP setting on Nikon scope to record images
-
{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table -->
+
Results:
-
|-valign="top"
+
* Saw some cells with very bright signal, some dim in 0 μg/mL dox well
-
| <u>Reagent</u> || <u>Volume</u>
+
* Signal appeared to be dimmer overall for all others.
-
|-
+
-
| reagent 1 || # μL
+
-
|-
+
-
| reagent 2 || #
+
-
|-
+
-
| reagent 3 || #
+
-
|-
+
-
| reagent 4 || #
+
-
|-
+
-
| dH<sub>2</sub>O || #
+
-
|-
+
-
| &nbsp; || # μL
+
-
|}
+
-
--> Reaction conditions
 

Current revision

Pc-TF Genomics Main project page
Previous entry      Next entry


06/13/13

  • Gal4-EED/luc cells IFC - secondary stain & imaging



Gal4-EED/luc cells IFC

  • Samples: first row of wells in the 24-well plate.
  • Keep all others under PBS
  1. 0 μg/mL dox
  2. 0.1
  3. 0.2
  4. 0.5
  5. 1.0
  6. 2.0
  • Follow standard protocol; wash 3x with 1xPBS
  • Use 1:1000 dilution of anti-rabbit Alexa488 and 1:1000 Hoescht (in 15 μL Gentex horse serum blocking solution per well)
  • Incubate under parafilm "covers" for one hour at room temperature in light-tight box with wet paper towels
  • Use GFP setting on Nikon scope to record images

Results:

  • Saw some cells with very bright signal, some dim in 0 μg/mL dox well
  • Signal appeared to be dimmer overall for all others.




Personal tools