TRNT1 Overexpression Lysate

Images

 
Western Blot: TRNT1 Overexpression Lysate (Adult Normal) [NBL1-17327] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for TRNT1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

TRNT1 Overexpression Lysate Summary

Description

TRNT1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC203043

Accession#: NM_016000

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
TRNT1

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.

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 TRNT1 Overexpression Lysate

  • CCA1
  • CCA-adding
  • CGI-47
  • EC 2.7.7.72
  • mitochondrial CCA-adding tRNA-nucleotidyltransferase
  • mt CCA-adding enzyme
  • mt tRNA adenylyltransferase
  • mt tRNA CCA-diphosphorylase
  • mt tRNA CCA-pyrophosphorylase
  • MtCCA
  • tRNA nucleotidyl transferase, CCA-adding, 1
  • tRNA-nucleotidyltransferase 1, mitochondrial

Background

The CCA-adding enzyme TRNT1 (EC 2.7.7.25) is an essential enzyme that catalyzes the addition of the CCA terminus to the 3-prime end of tRNA precursors. This reaction is a fundamental prerequisite for mature tRNAs to become aminoacylated and to participate in protein biosynthesis (Lizano et al., 2007 (PubMed 17204286)).(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.

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Video Protocols

WB Video Protocol

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Bioinformatics

Gene Symbol TRNT1