Intersectin 1 Overexpression Lysate

Western Blot: Intersectin 1 Overexpression Lysate (Adult Normal) [NBP2-10857] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Intersectin 1.

• Reactivity: Hu
• Applications: WB

Intersectin 1 Overexpression Lysate Summary

• Description:

  Intersectin 1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC221701
  
  Accession#: NM_003024
  
  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: ITSN1

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: 195.2 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 Intersectin 1 Overexpression Lysate

• 11
• intersectin 1 (SH3 domain protein)
• intersectin short variant 12
• intersectin-1
• ITSN
• SH3 domain protein-1A
• SH3 domain-containing protein 1A
• Src homology 3 domain-containing protein

Background

Intersectin 1 is an adapter protein that may provide an indirect link between the endocytic membrane traffic and the actin assembly machinery. It may regulate the formation of clathrin-coated vesicles. Isoform 1 could be involved in brain-specific synaptic vesicle recycling. Intersectin 1 is overexpressed in brain from Down syndrome foetuses suggesting a gene dosage-dependent contribution to the abnormalities of Down syndrome. Intersectin 1 interacts with dynamin, CDC42, SNAP25 and SNAP23. It clusters several dynamin in a manner that is regulated by alternative splicing. Intersectin 1 also binds clathrin-associated proteins and other components of the endocytic machinery, such as SPIN90, EPS15, EPN1, EPN2 and STN2. Adapter protein that may provide indirect link between the endocytic membrane traffic and the actin assembly machinery. May regulate the formation of clathrin-coated vesicles. Isoform 1 could be involved in brain-specific synaptic vesicle recycling.

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: ITSN1