Human Recombinant Microtubule Associated Protein Tau (MAPT) Lyophilized

Properties

• Size: 200 μg
• Appearance: Lyophilized powder
• Formulation: Lyophilized from 1 mg/ml Tau in PBS, pH 7.4 solution, filtered through 0.2 μm pore size Whatman filter paper. Contact us for customized product form or formulation.
• Reconstitution: Reconstitute in desired buffer. It is recommended to add reducing agent such as DTT or 2-mercapto-ethanol
• Purification: Purified by ion-exchange chromatography.
• Purity: > 95% as determined by SDS-PAGE.
• Endotoxins: ND
• Storage and Stability: Once reconstituted, it is recommended to freeze aliquots at - 80°C for up to 12 months under sterile conditions. Avoid freeze-thaw cycles.
• Shipping: This product is shipped with dry ice in Europe, may vary elsewhere. Please inquire the shipping cost.
• Cite This Product: Human Recombinant Microtubule Associated Protein Tau lyophilized powder (BRINDx, Argentina, REF: BRX-2002).

Biological Descriptions

• Background: Promotes microtubule assembly and stability, and might be involved in the establishment and maintenance of neuronal polarity (PubMed:21985311).
  The C-terminus binds axonal microtubules while the N-terminus binds neural plasma membrane components, suggesting that tau functions as a linker protein between both (PubMed:21985311, PubMed:32961270).
  Axonal polarity is predetermined by TAU/MAPT localization (in the neuronal cell) in the domain of the cell body defined by the centrosome. The short isoforms allow plasticity of the cytoskeleton whereas the longer isoforms may preferentially play a role in its stabilization.
• Research area: Neuroscience. Neurogenerative disorders. Alzheimer’s disease.  Tauopathies.
• Cellular Localization: Mostly found in the axons of neurons, in the cytosol and in association with plasma membrane components (PubMed:10747907).
  Can be secreted; the secretion is dependent on protein unfolding and facilitated by the cargo receptor TMED10; it results in protein translocation from the cytoplasm into the ERGIC (endoplasmic reticulum-Golgi intermediate compartment) followed by vesicle entry and secretion (PubMed:32272059).
• Synonyms: MAPTL, MTBT1, TAU. Data Base: Gene ID: 4137 [NCBI].
• Accession: #P10636.

Product Data

SDS PAGE of Tau protein monomer (left lane) along with a molecular weight marker (right lane)

FPLC. Tau protein Chromatrogram purification . Elution of the protein by performing a NaCl concentration gradient up to 60%. The peak with the highest absorbance corresponds to the purified protein.

Applications

Tau aggregates seeding capability and citotoxicity

 (A) Effect of doxycycline on the seeding capability of pre-aggregated tau species. Pre-incubated tau species were obtained by incubating a tau:heparin solution (tau 22 μM; 0.2 mg/ml heparin) 24 h at 37°C under orbital agitation. 25 μl aliquots of this solution were used as seeds and added to fresh monomeric tau (22 μM) in the absence or presence of doxycycline 100 μM and incubated 0 h, 48 h and 168 h at 37°C under orbital agitation. (B) SH-SY5Y cells were treated with aggregated species of tau incubated in the presence of heparin, with or without doxycycline, and harvested after 16 h of incubation. Viability was measured using an MTT assay and expressed as a percentage relative to the untreated cells. Statistical analyses for both experiments were performed using the ANOVA test. n = 3 ns: not significant. ****p ≤ 0.0001. Error bars represent SD.

References

1. Yoshida, H., and Goedert, M. (2012) Phosphorylation of microtubule-associated protein tau by AMPK-related kinases. J. Neurochem. 120, 165–76
2. Sandberg, A., Ling, H., Gearing, M., Dombroski, B., Cantwell, L., R’Bibo, L., Levey, A., Schellenberg, G. D., Hardy, J., Wood, N., Fernius, J., Nyström, S., Svensson, S., Thor, S., Hammarström, P., Revesz, T., and Mok, K. Y. (2020) Fibrillation and molecular characteristics are coherent with clinical and pathological features of 4-repeat tauopathy caused by MAPT variant G273R. Neurobiol. Dis. 146, 105079
3. Medina, L., González-Lizárraga, F., Dominguez-Meijide, A., Ploper, D., Parrales, V., Sequeira, S., Cima-Omori, M.-S., Zweckstetter, M., Del Bel, E., Michel, P. P., Outeiro, T. F., Raisman-Vozari, R., Chehín, R., and Socias, S. B. (2021) Doxycycline Interferes With Tau Aggregation and Reduces Its Neuronal Toxicity. Front. Aging Neurosci.