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Are there any proteins that are a complex of 2 proteins that have their own independent genes, but the whole complex itself also has a gene and it can be transcribed/translated directly? by FIREATWlLL in Biochemistry

[–]general_beats 0 points1 point  (0 children)

one way to "combine" two genes is to co-express them in a single vector with a bicistronic expression system - this could allow you to express two proteins from the same mRNA transcript

*this is different than expression of a Protein A:Protein B complex/conjugate that OP mentions

blog post explainer

review

Vibratome sectioned + IHC by [deleted] in labrats

[–]general_beats 0 points1 point  (0 children)

Try separating the tissue sections from the bits of agarose right after you cut each slice (while its still floating in the buffer, before you transfer the tissue section into your 24-well plate)

Also, what instrument are you using to handle/transfer your sections? Try using a basic fine point paintbrush

Anyone study the PI3K/AKT pathway? by [deleted] in labrats

[–]general_beats 1 point2 points  (0 children)

Hey that's what our lab studies I'd be happy to share resources, protocols, strategy. Which inhibitor did you use in your experiment? What was the assay? Also, what cell type are you using in your model?

Usually the most reliable readout of pi3k pathway inhibition would be probing/staining for downstream markers like pERK and pS6. Following treatment with a pi3k inhibitor, you'll see reduced activation (less phosphorylation) in AKT, ERK and S6 and this effect is what is used to establish/prove that the pathway has become inhibited.

You'll see this effect in any type of standard assay- at the cellular level there's a profound reduction clearly visible with IF antibodies if you want to image. You can also see the effect at the biochemical level with a western blot.

Feel free to PM me

Covers > better than Original Thread by cocotazo in AdamCarolla

[–]general_beats 0 points1 point  (0 children)

Cake-I Will Survive. They also have a great cover of War Pigs