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MCM3 Overexpression Lysate

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Western Blot: MCM3 Overexpression Lysate (Adult Normal) [NBL1-12951] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for MCM3.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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MCM3 Overexpression Lysate Summary

Description

MCM3 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC200208

Accession#: NM_002388

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
MCM3

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
90.8 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 MCM3 Overexpression Lysate

  • cervical cancer proto-oncogene 5
  • DNA polymerase alpha holoenzyme-associated protein P1
  • DNA replication factor MCM3
  • DNA replication licensing factor MCM3
  • EC 3.6.4.12
  • HCC5
  • hRlf beta subunit
  • MCM3 minichromosome maintenance deficient 3 (S. cerevisiae)
  • MCM3 minichromosome maintenance deficient 3
  • MGC1157
  • minichromosome maintenance complex component 3
  • minichromosome maintenance deficient (S. cerevisiae) 3
  • minichromosome maintenance deficient 3
  • P1.h
  • p102
  • P1-MCM3
  • replication licensing factor, beta subunit
  • RLF subunit beta
  • RLFB

Background

The protein encoded by this gene is one of the highly conserved mini-chromosome maintenance proteins (MCM) that are involved in the initiation of eukaryotic genome replication. The hexameric protein complex formed by MCM proteins is a key component of the pre-replication complex (pre_RC) and may be involved in the formation of replication forks and in the recruitment of other DNA replication related proteins. This protein is a subunit of the protein complex that consists of MCM2-7. It has been shown to interact directly with MCM5/CDC46. This protein also interacts with, and thus is acetlyated by MCM3AP, a chromatin-associated acetyltransferase. The acetylation of this protein inhibits the initiation of DNA replication and cell cycle progression. [provided by RefSeq]. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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 MCM3