In situ hybridization was carried out as described previously (He and Soriano, 2013 (link)). Pten antisense RNA probes was generated using primers: 5-TTACAGTTGGGCCCCGTACA-3 and 5- TAAAACACCCACACAATGACAAGA-3. For immunohistochemistry, samples were sectioned at 10μm and subjected to standard protocols using primary antibodies to anti-phospho-Akt (1:25, Cell Signaling Technology 9271), anti-Neurofilament-L (1:100, Cell Signaling Technology 2837), HRP-conjugated secondary antibody (1:100, Cell Signaling Technology 7074), anti-Sp7(1:200, Abcam ab94744) and AP-conjugated secondary antibody (1:100, Novus Biologicals, NB7157).)
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In Situ Hybridization and Immunohistochemistry
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Anti antibodies
Antibody
Biologicals
Immunohistochemistry
In situ hybridization
Neurofilament l
Novus
Primers
Pten
Rna probes
Corresponding Organization : Tulane University
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Protocol cited in 2 other protocols
Variable analysis
independent variables
- Pten antisense RNA probe generation using the specified primers
- In situ hybridization of Pten
- Phosphorylation of Akt
- Neurofilament-L expression
- Sp7 expression
- Sectioning of samples at 10μm
- Use of standard immunohistochemistry protocols
- Use of primary antibodies against phospho-Akt, Neurofilament-L, and Sp7
- Use of HRP-conjugated and AP-conjugated secondary antibodies
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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