NM23-H1 Overexpression Lysate

Western Blot: nm23-H1 Overexpression Lysate (Native) [NBP2-04154] - nm23-H1 Overexpression Lysate (Native) Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Nm23A.

• Reactivity: Hu
• Applications: WB

NM23-H1 Overexpression Lysate Summary

• Description:

  NM23-H1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC201731
  
  Accession#: NM_000269
  
  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: NME1

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: 17 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 NM23-H1 Overexpression Lysate

• EC 2.7.4.6
• GAAD
• Granzyme A-activated DNase
• Metastasis inhibition factor nm23
• NB
• NBS
• NDK A
• NDKA
• NDP kinase A
• NDPKA
• NDPK-A
• NM23A
• NM23AWD
• NM23H1
• NM23-H1
• NME1
• non-metastatic cells 1, protein (NM23A) expressed in
• nucleoside diphosphate kinase A
• Tumor metastatic process-associated protein

Background

The nm23 gene, a potential suppressor of metastasis, was originally identified by differential hybridization between two murine melanoma sub-lines, one with a high and the second with a low metastatic capacity. Highly metastatic sub-lines exhibit much lower levels of nm23 than less metastatic cells. Based on sequence analysis, nm23 appears highly related to nucleotide diphosphate kinases (NDP). In humans, NDP kinases A and B are identical to two isotypes of human nm23 homologs, namely nm23-H1 and -H2, respectively. nm23-H2 is identical in sequence to PuF, a transcription factor that binds to nucleasehypersensitive elements at positions 142 to 115 of the human c-Myc promotor.

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: NME1