11 In Situ Hybridization for RNA: Nonradioactive Probe: Oligo-DNA Probe
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- Nitrocellulose filter
- Vinyl bag for hybridization
- Gel stain Rotator
- Dryoven: Dry oven DS-44 I yamato, Japan
- Blocking solution
- 500 Ilg/ml normal goat IgG : Goat IgG, purified immunoglobulin (1-5256, Sigma)
- 5% BSA (98-99% pure): albumins, bovine (98-99% alb) (A7030, Sigma)
- 100 Ilg/ml yeast tRNA: RNA transfer, type X from Bakers
yeast (R-9001, Sigma)
- 100 Ilg/ml salmon sperm DNA: salmon testes DNA for hybridization (D-7656, Sigma)
- NaN3
- PBS
- 1st antibody
- mouse anti-T-T IgG (1: 200)
- 5% BSA (98-99% pure) : albumins,bovine (98-99% alb)
- 100 Ilg/ml yeast tRNA : RNA transfer, type X from Bakers
yeast
- 100 Ilg/ml salmon sperm DNA: salmon testes DNA for hybridization
- 0.05% NaN3
- PBS
- 2nd antibody
- HRP-goat anti-mouse IgG
- 5% BSA (98-99% pure) : albumins, bovine (98-99% alb)
- 100 Ilg/ml yeast tRNA : RNA transfer,type X from Bakers
yeast
- 100 Ilg/ml salmon sperm DNA: salmon testes DNA for hybridization
- 0.05% NaN3
- PBS
- DAB/H20 2/Co/Ni solution
- 1M PB : 1M Na2HP04 and 1M NaH2P04 (with DDW)
- 1% CoClz
- 1% NiS04-NH4S04
- 31% H20 Z
- DAB
for immunohistochemical
detection of
T-T dimerized
DNA on a
nitrocellulose
filter