EGLN3/PHD3 Overexpression Lysate

Novus Biologicals | Catalog # NBL1-10156

Novus Biologicals
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for EGLN3/PHD3 Overexpression Lysate

EGLN3/PHD3 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC210319

Accession#: NM_022073

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.
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Product Specifications

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

27.1 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.

Type

Overexpression

Scientific Data Images for EGLN3/PHD3 Overexpression Lysate

Western Blot: EGLN3/PHD3 Overexpression Lysate [NBL1-10156]

Western Blot: EGLN3/PHD3 Overexpression Lysate [NBL1-10156]

Western Blot: PHD3/HIF Prolyl Hydroxylase 3 Overexpression Lysate (Adult Normal) [NBL1-10156] - Left - Empty vector transfected control cell lysate (HEK293 cell lysate); Right - Over-expression Lysate for HIF Prolyl Hydroxylase 3.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: EGLN3/PHD3

HIF prolyl hydroxylase 3 (HPH-3; PHD3; EGLN3) catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins, hydroxylates HIF-1 alpha at Pro-564, and also hydroxylates HIF-2 alpha. PHD3 functions as a cellular oxygen sensor and, under normoxic conditions, targets HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitylation complex. EGLN3 may play a role in cell growth regulation in muscle cells and in apoptosis in neuronal tissue. HIF prolyl hydroxylase 3 promotes cell death through a caspase-dependent mechanism.

Long Name

Egl Nine Homolog 3/Prolyl Hydroxylase Domain-containing Protein 3

Alternate Names

HIFPH3, PHD3

Gene Symbol

EGLN3

Additional EGLN3/PHD3 Products

Product Documents for EGLN3/PHD3 Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for EGLN3/PHD3 Overexpression Lysate

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.

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.

Citations for EGLN3/PHD3 Overexpression Lysate

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: HEK293T Whole cell lysate transfected with EGLN3 plasmid
    Species: Human
    Verified Customer | Posted 12/07/2015
    EGLN3/PHD3 Overexpression Lysate NBL1-10156

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Associated Pathways

HIF Enhancer Pathways HIF Enhancer Pathway Thumbnail