Applications: Suitable for use in Western Blot and Immunohistochemistry. Other applications have not been tested. Recommended Dilution: Western Blot: 5ng/ml. Bovine brain microtubule proteins purified by two cycles of assembly and disassembly (9) are dissolved in SDS-PAGE sample buffer. 5ug of the microtuble preparation per lane is loaded onto a 4% to 20% SDS-PAGE gradient gel along side molecular weight markers (14.3-200kD). After separation by electrophoresis, the proteins are blotted onto nitrocellulose. Tau is detected as a series of 5 bands (52-68kD). Immunohistochemistry: 5ug/ml. Stains axons in tissue primarily, however in culture Tau expression is not restricted to just axons. Dephosphorylation with alkaline phosphatase is recommended for staining neurofibrillary tangles in Alzheimers brain tissue with T1037. This treatment changes the staining pattern of T1037 to include cell bodies, dendrites and axons of neurons. In untreated samples, T1037 stains axons only. Immunofluorescence:1000 detected Tau in mouse primary neurons. (Basnet, N., et al. (2018). Nat. Cell Biol. 20(10), 1172-1180. Optimal dilutions to be determined by the researcher. Positive Control: Alzheimer's brain tissue (dephosphorylation with alkaline phosphatase is recommended for staining neurofibrillary tangles in Alzheimers brain tissue) or human T98G glioblastoma cells Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
