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Tau Overexpression Lysate

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Western Blot: Tau Overexpression Lysate (Adult Normal) [NBP2-06994] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Tau.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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Tau Overexpression Lysate Summary

Description

Tau Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC213364

Accession#: NM_016841

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Gene
MAPT

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Theoretical MW
36.6 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for Tau Overexpression Lysate

  • DDPAC
  • FLJ31424
  • FTDP-17
  • G protein beta1/gamma2 subunit-interacting factor 1
  • MAPT
  • MGC138549
  • microtubule-associated protein tau
  • MSTD
  • MSTDMAPTL
  • MTBT1
  • MTBT1Neurofibrillary tangle protein
  • MTBT2
  • Neurofibrillary tangle protein
  • PHF-tau
  • PPND
  • Tau
  • TAUPaired helical filament-tau

Background

Tau is a microtubule binding protein that promotes microtubule assembly and stability. Tau is found to be the major component of the paired helical filaments (PHFs) found in the brains of patients with Alzheimer disease (AD) (1,2). Tau is hyperphosphorylated in PHFs, and specific phosphorylation sites have been implicated in the loss of Tau's association with the membrane cortex during AD disease state, including Ser 199/202, Thr 231, and Ser 393/404 (3). Glycogen synthase kinase-3, or GSK-3, phosphorylates Tau on Ser 396 (4).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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Product General Protocols

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Video Protocols

WB Video Protocol

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Alzheimer’s Disease (AD) is a neurodegenerative disorder that is characterized by an abundance of the beta-amyloid peptide in the brain.  When AD was first discovered, it was determined that beta-amyloid was produced as a result of the prote...  Read full blog post.

Tau - A microtubule associated protein as a biomarker for Alzheimer's disease
The tau protein is a microtubule associated protein found mostly in neuronal cells where it regulates the stability of axonal microtubules as well as kinesin-dependent transport. Tau is relevant in the study of various neurological disorders as ab...  Read full blog post.

PINK1: All work and no fun
The protein PINK1 is a mitochondrial-located serine/threonine kinase (PTK) that maintains organelle function and integrity. It not only protects organelles from cellular stress, but it also uses the selective auto-phagocytosis process for cleaning and...  Read full blog post.

Using Amyloid beta peptides in Alzheimer's Disease Immunization
Amyloid beta (AB) peptide has a central role in the neurodegeneration of Alzheimer's disease (AD). Immunization of AD transgenic mice with AB-42 peptide reduces both the spatial memory impairments and AD-like neuropathologic changes.Therapeutic im...  Read full blog post.

New Study Links Tau Mutations to Microglial Immune Response
Tau proteins are abundant in the axons of neurons in the central nervous system (CNS), and play a key role in microtubule formation and stabilization. Antibody studies have identified six tau isoforms, all produced by alternative mRNA splicing of the ...  Read full blog post.

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Bioinformatics

Gene Symbol MAPT