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RENT1/UPF1/hUPF1 Overexpression Lysate

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Western Blot: hUPF1 Overexpression Lysate (Adult Normal) [NBP2-10854] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for hUPF1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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RENT1/UPF1/hUPF1 Overexpression Lysate Summary

Description

RENT1/UPF1/hUPF1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC208018

Accession#: NM_002911

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
UPF1

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
122.9 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 RENT1/UPF1/hUPF1 Overexpression Lysate

  • ATP-dependent helicase RENT1
  • EC 3.6.1
  • EC 3.6.4.-
  • FLJ43809
  • HUPF1
  • KIAA0221FLJ46894
  • Nonsense mRNA reducing factor 1
  • NORF1
  • NORF1delta helicase
  • pNORF1
  • regulator of nonsense transcripts 1
  • RENT1
  • RENT1UP Frameshift 1
  • UPF1 regulator of nonsense transcripts homolog (yeast)
  • UPF1
  • up-frameshift mutation 1 homolog
  • Up-frameshift suppressor 1 homolog
  • yeast Upf1p homolog

Background

hUPF1 encodes a protein that is part of a post-splicing multiprotein complex involved in both mRNA nuclear export and mRNA surveillance. mRNA surveillance detects exported mRNAs with truncated open reading frames and initiates nonsense-mediated mRNA decay (NMD). When translation ends upstream from the last exon-exon junction, this triggers NMD to degrade mRNAs containing premature stop codons. This protein is located only in the cytoplasm. When translation ends, it interacts with the protein that is a functional homolog of yeast Upf2p to trigger mRNA decapping. Use of multiple polyadenylation sites has been noted for this gene. [provided by RefSeq]

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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Bioinformatics

Gene Symbol UPF1