PTTG1 Overexpression Lysate

Western Blot: Securin Overexpression Lysate (Native) [NBP2-07897] - Securin Overexpression Lysate (Native) Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PTTG1.

• Reactivity: Hu
• Applications: WB

PTTG1 Overexpression Lysate Summary

• Description:

  PTTG1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC211038
  
  Accession#: NM_004219
  
  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: PTTG1

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: 21.8 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 PTTG1 Overexpression Lysate

• EAP1MGC126883
• ESP1-associated protein 1
• Esp1-associated protein
• hPTTG
• pituitary tumor-transforming 1
• Pituitary tumor-transforming gene 1 protein
• PTTGMGC138276
• securin
• Tumor-transforming protein 1
• TUTR1HPTTG

Background

The product of the oncogene PTTG, pituitary tumor transforming gene, is a human homolog of the anaphase-inhibitor vertebrate protein, securin. PTTG is a 22 kDa protein that contains a basic amino-terminal domain and an acidic carboxy-terminal domain, which acts as a transactivation domain when fused to a heterologous DNA binding domain. Human PTTG is overexpressed in Jurkat and is also detected in human thymus, testis, and placenta. PTTG is mainly expressed in the cytoplasm and is also partially localized to the nucleus. Vertebrate PTTG regulates the separin Esp1, which promotes chromatid separation, to overcome the cohesive forces that hold sister chromatids together. This regulatory function of PTTG suggests that defective regulation of cohesion may contribute to cancer by promoting chromosome instability. Although vertebrate PTTG shares cell-cycle functions with its yeast securin counterparts Pds1p and Cut2, none share sequence homology.

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: PTTG1