i am working on STZ induced sporadic Alzheimer’s Disease model. i was wondering about the best protein to assess by western blot after 6 weeks of induction considering that i have limited resources. Moreover, what is the best source to purchase the antibodies?
thanks in advance
I assume you wish to test whether your experiment caused recution of the IR protein levels as well as it's activity. For this you will actually need three antibodies. The first is agains the IR beta-subunit and the two otheres should be to the phosphorylated form pTyr1162/1163 and the pTyr1158. Nevertheless, if you are able it would be better to use ELISA method as you can quantify your protein in a much more acurate manner. As for example in two papers we published
Grünblatt E, Bartl J, Iuhos DI, Knezovic A, Trkulja V, Riederer P, Walitza S,
Salkovic-Petrisic M. Characterization of cognitive deficits in spontaneously
hypertensive rats, accompanied by brain insulin receptor dysfunction. J Mol
Psychiatry. 2015 Jun 4;3(1):6. doi: 10.1186/s40303-015-0012-6. eCollection 2015.
PubMed PMID: 26110057; PubMed Central PMCID: PMC4479234.
https://www.researchgate.net/publication/279164540_Characterization_of_cognitive_deficits_in_spontaneously_hypertensive_rats_accompanied_by_brain_insulin_receptor_dysfunction
Grünblatt E, Salkovic-Petrisic M, Osmanovic J, Riederer P, Hoyer S. Brain
insulin system dysfunction in streptozotocin intracerebroventricularly treated
rats generates hyperphosphorylated tau protein. J Neurochem. 2007
May;101(3):757-70. PubMed PMID: 17448147.
https://www.researchgate.net/publication/6380983_Brain_insulin_system_dysfunction_in_streptozotocin_intracerebroventricularly_treated_rats_generates_hyperphosphorylated_tau_protein_J_Neurochem
Article Characterization of cognitive deficits in spontaneously hype...
Article Brain insulin system dysfunction in streptozotocin intracere...
IR, IRS, pAKT etc would should tell you the insulin status. If sample size is small can do the ELISA.
Thanks for the replies. So can i measure just IR and PI3K. If so do i need both unphosphorylated and phosphorylated forms or could it be just the phosphorylated form?
Measuring only the IR will only give you partial information which in some cases is already enough. But since there is compensatory mechanism of the IR, even if the protein is low the activity might still be high. That's why you need also to measure in this case also the phosphorylated form, as this will give you an indirect measure of the IR activity levels.
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