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HIF-3 alpha Overexpression Lysate

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Western Blot: HIF-3 alpha Overexpression Lysate (Adult Normal) [NBP2-05309] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for HIF-3 alpha.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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HIF-3 alpha Overexpression Lysate Summary

Description

HIF-3 alpha Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC210255

Accession#: NM_152795

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
HIF3A

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
72.3 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 HIF-3 alpha Overexpression Lysate

  • Basic-helix-loop-helix-PAS protein MOP7
  • BHLHe17
  • bHLHe17HIF-3A2
  • Class E basic helix-loop-helix protein 17
  • HIF-3 alpha
  • HIF3A
  • HIF-3A4
  • HIF-3-alpha
  • HIF3-alpha
  • hypoxia inducible factor 3, alpha subunit
  • hypoxia-inducible factor 3-alpha
  • hypoxia-inducible factor-3 alpha 4
  • Inhibitory PAS domain protein
  • IPAS
  • IPASHIF3-alpha-1
  • Member of PAS protein 7
  • MOP7
  • MOP7PAS domain-containing protein 7
  • PASD7
  • PASD7HIF-3A

Background

Hypoxia-inducible factor (HIF) is one of the most important factors in the cellular response to hypoxia, by transcriptionally activating genes encoding proteins that mediate adaptive responses to reduced oxygen availability. HIF is a heterodimer consisting of one of three subunits, HIF1 alpha, HIF2 alpha, or HIF3 alpha. HIF target genes play critical roles in metabolism, angiogenesis, cell proliferation and cell survival. HIF3 alpha protein is one of several alpha/beta-subunit heterodimeric transcription factors that regulate many adaptive responses to low oxygen tension (hypoxia). The alpha 3 subunit lacks the transactivation domain found in factors containing either the alpha 1 or alpha 2 subunits. HIF3 alpha may be a marker for tumor growth and angiogenesis.

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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Research Areas for HIF-3 alpha Lysate (NBP2-05309)

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Blogs on HIF-3 alpha.

HIF-2 alpha: HIF1A's Homologue with Similar and Divergent Functions
HIF-2 alpha is a member of the heterodimeric hypoxia-inducible factors/HIFs family (HIF-1, HIF-2, and HIF-3) which contains a common beta subunit but differ in their alpha subunits. Also called as EPAS1 or Mop2, HIF-2 alpha regulates cellular adapt...  Read full blog post.

HIF-3 alpha: a versatile target with hypoxia dependent and independent functions
By: Subhash GangarHIF-3 alpha (hypoxia-inducible factor 3-alpha/ HIF3A) represents an isoform of HIF-alpha subunits which heterodimerize with stable beta subunit (HIF-beta) for the regulation of HIF target genes through binding to hypoxia respon...  Read full blog post.

HIF Prolyl Hydroxylase 2: an important Oxygen Sensor Protein
Prolyl hydroxylase domain (PHD) proteins, including PHD1, PHD2, and PHD3, mediate oxygen-dependent degradation of hypoxia-inducible factor (HIF) alpha subunits. Suppression of PHD enzymes leads to stabilization of HIFs and offers a potential treatment...  Read full blog post.

HIF Antibodies: Beyond HIF-1 alpha
The hypoxia inducible factors are a family of heterodimeric transcription factors which are activated in response to lowered oxygen levels, or hypoxia. Although it may seem that HIF-1 alpha receives all the attention, other HIF antibodies, such as the...  Read full blog post.

A Role for HIF-1 alpha Antibody in Renal Research
The Hypoxia Inducible Factors (HIFs) are a family of mammalian transcription factors which are expressed in response to low cellular oxygen concentrations (hypoxia). Three human hypoxia inducible factors have been identified, HIF-1, HIF-2 and HIF-3, e...  Read full blog post.

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Bioinformatics

Gene Symbol HIF3A