SIX5 Overexpression Lysate

Western Blot: Six5 Overexpression Lysate (Adult Normal) [NBP2-05043] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Six5.

• Reactivity: Hu
• Applications: WB

SIX5 Overexpression Lysate Summary

• Description:

  SIX5 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC211844
  
  Accession#: NM_175875
  
  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: SIX5

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: 74.4 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 SIX5 Overexpression Lysate

• BOR2
• DM locus-associated homeodomain protein
• DMAHPsine oculis homeobox homolog 5 (Drosophila)
• dystrophia myotonica-associated homeodomain protein
• homeobox protein SIX5
• sine oculis homeobox (Drosophila) homolog 5
• Sine oculis homeobox homolog 5
• SIX homeobox 5

Background

Six5 (homeobox protein SIX5), also known as SIX5, BOR2 or DMAHP (DM locus-associated homeodomain protein), is a transcription factor that is expressed in various structures of the adult eye. Localized to the cytoplasm in early development and to the nucleus in the later stages of development,Six5 is involved in regulation of organogenesis and in maintenance of retinal formation. Six5 is able to bind the 5'-TCA[AG][AG]TTNC-3' DNA sequence found in the myogenin and IGFBP5 promoters and, through this binding, can control transcription of the associated mRNA. Six5 is regulated via association with DACH1 (dachshund homolog 1) and is co-activated by the EYA (eyes absent) proteins. Defects in the gene encoding Six5 are the cause of branchiooto- renal syndrome type 2 (BOR2), an autosomal disorder characterized by hearing loss, a deep overbite and myopia. Two isoforms exist due to alternative splicing events.

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: SIX5