Etchevers:Notebook/Genomics of hNCC/2009/04/14: Difference between revisions
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== | ==Fed cells== | ||
The R1066 cephalic cells (that I mistakenly labelled R1113 for the mycoplasma test that Stephane will do) are doing alright but not wildly proliferating, either. | |||
The rest of the Rich media was a little murky - I filter-sterilized it without looking to see if it was yeast. Probably was. Used the Stericup and there are probably no nutrients left. Sigh. | |||
Stephane's EC cultures are either mostly dead, few cells or exploding with confluent cells. He only kept the last one, will test the mycoplasma as well, and split it 1:2 into two new wells of a 6-well plate. Actually the one with dead cells I think he rinsed and then re-seeded into a 12- or 24-well plate (1 well). | |||
Immunocytofluorescence. | |||
Finally used 1:100 anti-tubulin beta 3 (IgG1) and 1:100 anti-tyrosinase (IgG2a), incubated 2h at RT after blocking 1h in PBT/1x BR that had been filtered as I had left it to melt over the whole weekend and things might grow in it. | |||
Then 1.5 hours of 1:100 anti-IgG1 (633) and 1:100 anti-IgG2a (555) before extensive rinsing and mounting in Prolong Gold with DAPI. Keep in fridge until Friday. | |||
*'''[[User:Etchevers|Heather]] 12:05, 14 April 2009 (EDT)''': | |||
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Revision as of 09:05, 14 April 2009
 Genomics of human neural crest cells
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Fed cellsThe R1066 cephalic cells (that I mistakenly labelled R1113 for the mycoplasma test that Stephane will do) are doing alright but not wildly proliferating, either. The rest of the Rich media was a little murky - I filter-sterilized it without looking to see if it was yeast. Probably was. Used the Stericup and there are probably no nutrients left. Sigh. Stephane's EC cultures are either mostly dead, few cells or exploding with confluent cells. He only kept the last one, will test the mycoplasma as well, and split it 1:2 into two new wells of a 6-well plate. Actually the one with dead cells I think he rinsed and then re-seeded into a 12- or 24-well plate (1 well). Immunocytofluorescence. Finally used 1:100 anti-tubulin beta 3 (IgG1) and 1:100 anti-tyrosinase (IgG2a), incubated 2h at RT after blocking 1h in PBT/1x BR that had been filtered as I had left it to melt over the whole weekend and things might grow in it. Then 1.5 hours of 1:100 anti-IgG1 (633) and 1:100 anti-IgG2a (555) before extensive rinsing and mounting in Prolong Gold with DAPI. Keep in fridge until Friday.
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