APBA3/Mint-3 Overexpression Lysate

Western Blot: APBA3/Mint-3 Overexpression Lysate [NBP2-10542] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Mint 3.

• Reactivity: Hu
• Applications: WB

APBA3/Mint-3 Overexpression Lysate Summary

• Description:

  APBA3/Mint-3 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC210256
  
  Accession#: NM_004886
  
  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: APBA3

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: 61.3 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 APBA3/Mint-3 Overexpression Lysate

• Adapter protein X11gamma
• amyloid beta (A4) precursor protein-binding, family A, member 3 (X11-like 2)
• amyloid beta (A4) precursor protein-binding, family A, member 3
• APBA3
• MGC:15815
• MINT3
• mint-3
• Neuronal Munc18-1-interacting protein 3
• Neuron-specific X11L2 protein
• phosphotyrosine-binding/-interacting domain (PTB)-bearing protein
• X11L2
• X11L2amyloid beta A4 precursor protein-binding family A member 3
• X11-like 2 protein

Background

The munc-18 interacting (Mint) protein family is a group of evolutionarily conserved adaptor proteins that function in membrane transport and organization. In mammals, there exist three mint isoforms, Mint 1, 2, and 3. Although there is little amino acid sequence conservation in the amino-terminal half, the carboxy-terminal half of these proteins is highly conserved. Within this conserved portion there exists a phosphotyrosine-binding (PTB) and a PSD-95/DLG-A/ZO-1 (PDZ) domain, which function as protein interaction modules. Mint 1 and 2 appear to be expressed exclusively in the brain and are found to bind to munc-18, an essential component of the synaptic vesicle fusion machinery. Mint 3 is ubiquitously expressed in all tissues and is expressed at the lowest levels in the brain and testis. Studies show that Mint 3 does not interact with munc-18. Mint 3 has been found to interact with the Alzheimer®s Disease-related amyloid precursor protein (APP) and does so through its PTB and PDZ domains. It has been suggested that Mint 3 links APP to other transport machinery components, thereby regulating it transport, endocytosis, and metabolism. Abnormal APP metabolism has been shown to be the cause of an early-onset type of Alzheimer®s disease.

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: APBA3