Kua-UEV Overexpression Lysate

Western Blot: TMEM189 Overexpression Lysate (Adult Normal) [NBP2-04652] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for TMEM189.

• Reactivity: Hu
• Applications: WB

Kua-UEV Overexpression Lysate Summary

• Description:

  Kua-UEV Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC223133
  
  Accession#: NM_199203
  
  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: TMEM189-UBE2V1

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: 42 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 Kua-UEV Overexpression Lysate

• CROC1
• CROC-1
• CROC1B
• CROC-1B
• Kua-UEV
• TMEM189-UBE2V1 fusion protein
• TMEM189-UBE2V1 readthrough transcript
• TMEM189-UBE2V1 readthrough
• TRAF6-regulated IKK activator 1 beta Uev1A
• transmembrane protein 189-ubiquitin-conjugating enzyme E2 variant 1read-through
• UBE2V
• UEV1
• UEV-1

Background

The Kua-UEV mRNA is an infrequent but naturally occurring co-transcribed product of the neighboring Kua and UBE2V1 genes. Ubiquitin-conjugating E2 enzyme variant proteins constitute a distinct subfamily within the E2 protein family. They have sequence similarity to other ubiquitin-conjugating enzymes but lack the conserved cysteine residue that is critical for the catalytic activity of E2s. Two alternative transcripts encoding different isoforms have been described. The proteins produced by these transcripts have UEV1 B domains but the proteins are localized to the cytoplasm rather than to the nucleus. The significance of these co-transcribed mRNAs and the function of their protein products have not yet been determined.

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: TMEM189-UBE2V1