NKX6.1 Overexpression Lysate

Western Blot: Nkx6.1 Overexpression Lysate (Adult Normal) [NBL1-13663] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Nkx6.1.

• Reactivity: Hu
• Applications: WB

NKX6.1 Overexpression Lysate Summary

• Description:

  NKX6.1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC224762
  
  Accession#: NM_006168
  
  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: NKX6-1

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: 37.7 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 NKX6.1 Overexpression Lysate

• Homeobox protein NK-6 homolog A
• NK homeo box, family 6, member A
• NK homeobox (Drosophila), family 6, A
• NK homeobox, family 6, A
• NK6 homeobox 1
• NK6 transcription factor homolog A
• NK6 transcription factor related, locus 1 (Drosophila)
• NK6 transcription factor related, locus 1
• NKX6.1
• NKX6-1
• NKX6A
• NKX6Ahomeobox protein Nkx-6.1

Background

In the pancreas, NKX6.1 is required for the development of beta cells and is a potent bifunctional transcription regulator that binds to AT-rich sequences within the promoter region of target genes Iype et al. (2004) [PubMed 15056733].[supplied by OMIM]

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.

Publications for NKX6.1 Lysate (NBL1-13663)(1)

Publication using NBL1-13663

• Wheeler E, Huang N, Bochukova EG et al. Genome-wide SNP and CNV analysis identifies common and low-frequency variants associated with severe early-onset obesity Nat Genet 2013-04-07 [PMID: 23563609]

Bioinformatics

• Gene Symbol: NKX6-1