beta-III Tubulin Overexpression Lysate

Western Blot: Tubulin Beta 3 Overexpression Lysate (Adult Normal) [NBL1-17440] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Tubulin Beta 3.

• Reactivity: Hu
• Applications: WB

beta-III Tubulin Overexpression Lysate Summary

• Description:

  beta-III Tubulin Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC200755
  
  Accession#: NM_006086
  
  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: TUBB3

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: 50.3 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 beta-III Tubulin Overexpression Lysate

• Beta3-tubulin
• beta-4
• betaIII Tubulin
• beta-III Tubulin
• beta-Tubulin III
• Class III beta-tubulin
• TUBB3
• TUBB4 CFEOM3A
• Tubulin Beta 3
• tubulin beta-3 chain
• Tubulin beta-4 chain
• Tubulin beta-4
• Tubulin beta-III
• tubulin, beta 3
• TUJ1 antigen
• TUJ-1 antigen

Background

Beta-III Tubulin is abundant in the central and peripheral nervous systems where it is prominently expressed during fetal and postnatal development. Beta III Tubulin is often referred to as TUJ1 because Tuj-1 is the clone name of the original monoclonal antibody. The Tuj1 clone is such a popular neuronal marker that Tuj-1 is now used as a synonym for beta III tubulin. The distribution of Tubulin Beta 3 is neuron-associated, exhibiting distinct temporospatial gradients according to the regional neuroepithelia of origin. However, transient expression of the Tubulin Beta 3 protein is also present in the subventricular zones of the CNS comprising putative neuronal- and/or glial precursor cells, as well as in Kulchitsky neuroendocrine cells of the fetal respiratory epithelium. This temporally restricted, potentially non-neuronal expression may have implications in the identification of presumptive neurons derived from embryonic stem cells. Mutation on beta-tubulin has been linked to anti-cancer drug resistance in lung cancers.

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.

Bioinformatics

• Gene Symbol: TUBB3