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XPB Overexpression Lysate

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Western Blot: XPB Overexpression Lysate (Adult Normal) [NBL1-10321] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for XPB.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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XPB Overexpression Lysate Summary

Description

XPB Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC202425

Accession#: NM_000122

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
ERCC3

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
89.1 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 XPB Overexpression Lysate

  • Basic transcription factor 2 89 kDa subunit
  • BTF2 p89
  • BTF2
  • DNA excision repair protein ERCC-3
  • DNA repair protein complementing XP-B cells
  • EC 3.6.1
  • EC 3.6.4.12
  • ERCC3
  • excision repair cross-complementing rodent repair deficiency, complementationgroup 3 (xeroderma pigmentosum group B complementing)
  • GTF2H
  • RAD25
  • TFIIH basal transcription factor complex 89 kDa subunit
  • TFIIH basal transcription factor complex helicase XPB subunit
  • TFIIH p89
  • TFIIH
  • Xeroderma pigmentosum group B-complementing protein
  • xeroderma pigmentosum, complementation group B
  • XPB
  • XPBC
  • XPBTFIIH 89 kDa subunit

Background

ERCC3 is a component of the core TFIIH basal transcription factor and functions as an ATP-dependent 3'-5' DNA helicase. ERCC3 is involved in nucleotide excision repair and is the cause of xeroderma pigmentosum complementation group B (XPB), also known as xeroderma pigmentosum II (XP2). Additionally, it has been found to be the cuase of Cockayne syndrome, and trichothiodystrophy (TTD). XPB is an autosomal recessive disease characterized by skin photosensitivity and a predisposition to skin cancer, and neurological abnormalities. Cockayne syndrome and TTD are similarly characterized by photosensitivity and neural abnormalities.

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

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Bioinformatics

Gene Symbol ERCC3