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POU3F2/OCT7 Overexpression Lysate

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Western Blot: OCT7 Overexpression Lysate (Adult Normal) [NBP2-10458] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for OCT7.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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POU3F2/OCT7 Overexpression Lysate Summary

Description

POU3F2/OCT7 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC205330

Accession#: NM_005604

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
POU3F2

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 (cat# 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
46.7 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 POU3F2/OCT7 Overexpression Lysate

  • Brain-2
  • Brain-specific homeobox/POU domain protein 2
  • BRN2
  • brn-2
  • BRN2brain-2
  • Nervous system-specific octamer-binding transcription factor N-Oct-3
  • Oct7
  • oct-7
  • OCT7POU domain class 3, transcription factor 2
  • Octamer-binding protein 7
  • Octamer-binding transcription factor 7
  • OTF7
  • OTF-7
  • POU class 3 homeobox 2
  • POU domain, class 3, transcription factor 2
  • POU3F2
  • POUF3

Background

POU3F2 belongs to a large family of transcription factors that bind to the octameric DNA sequence ATGCAAAT. Most of these proteins share a highly homologous region, referred to as the POU domain, that occurs in several mammalian transcription factors, including the octamer-binding proteins Oct1 (POU2F1; MIM 164175) and Oct2 (POU2F2; MIM 164176) and the pituitary protein Pit1 (PIT1; MIM 173110). Class III POU genes are expressed predominantly in the central nervous system (CNS). It is likely that CNS-specific transcription factors such as these play an important role in mammalian neurogenesis by regulating their diverse patterns of gene expression (Schreiber et al., 1993 [PubMed 8441633]; Atanasoski et al., 1995 [PubMed 7601453]).[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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Product General Protocols

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Video Protocols

WB Video Protocol

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Bioinformatics

Gene Symbol POU3F2