DOK6 Overexpression Lysate

Western Blot: DOK6 Overexpression Lysate (Adult Normal) [NBL1-09981] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for DOK6.

• Reactivity: Hu
• Applications: WB

DOK6 Overexpression Lysate Summary

• Description:

  DOK6 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC210487
  
  Accession#: NM_152721
  
  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: DOK6

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: 38.1 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 DOK6 Overexpression Lysate

• docking protein 5-like
• docking protein 6
• DOK5L
• Downstream of tyrosine kinase 6
• HsT3226
• MGC20785

Background

The downstream of kinase family (Dok1-6) are members of a class of "docking" proteins that include the tyrosine kinase substrates IRS-1 and Cas, which contain multiple tyrosine residues and putative SH2 binding sites. Dok-4, Dok-5 and Dok-6 are more similar to each other than to the other Dok family members, and may constitute a subfamily of the DOK genes. Dok-5 is a tyrosine kinase substrate that enhances c-Ret-dependent activation of mitogen-activated protein kinase (MAPK). Dok-5 transcript is abundant in muscle and increases during T cell activation. Dok-5 protein undergoes tyrosine phosphorylation in response to insulin and insulin-like growth factor-1. The gene encoding human Dok-5 maps to chromosomal location 20q13.2. Dok-6 is highly expressed in the developing central nervous system. It associates with Ret to transduce Ret-mediated processes such as axonal projection.

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: DOK6