NEK6 Overexpression Lysate

Western Blot: NEK6 Overexpression Lysate (Adult Normal) [NBP2-10144] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NEK6.

• Reactivity: Hu
• Applications: WB

NEK6 Overexpression Lysate Summary

• Description:

  NEK6 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC228277
  
  Accession#: NM_001166171
  
  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: NEK6

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: 39.7 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 NEK6 Overexpression Lysate

• EC 2.7.11.1
• Never in mitosis A-related kinase 6
• NIMA (never in mitosis gene a)-related kinase 6
• NimA-related protein kinase 6
• Protein kinase SID6-1512
• putative serine-threonine protein kinase
• serine/threonine-protein kinase Nek6
• SID6-1512

Background

NEK6 encodes a NEK type protein kinase, which contains a C-terminal NIMA-like catalytic domain. The protein is believed to regulate chromatin condensation in mammalian cells via phosphorylation of histones H1 and H3, and it has been shown to phosphorylate ribosomal S6 protein kinase (S6K1) and serum and glucocorticoid induced protein kinase (SGK1), which regulate diverse cellular processes. The protein also has been shown to bind NEK9, another NIMA-related kinase which regulates chromosome alignment and segregation. At least three mRNA transcripts have been identified, 1.6-, 2.6-, and 9.5-kb. The NEK6 gene is localized to 9q33-34, a region at which the loss of heterozygosity is associated with transitional cell carcinomas. NEK6 expression has been documented in many normal human tissues, including blood, brain, heart, liver, lung, skeletal muscle, spleen and testis. ESTs have been isolated from numerous tissue libraries, especially normal cervix and amnion and cancerous bone marrow.

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: NEK6