Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Primate

Predicted:

Orangutan (100%), Rabbit (100%), Rhesus Macaque (100%). Backed by our 100% Guarantee.

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Western Blot (Negative)

Label

Alexa Fluor 647 (Excitation = 650 nm, Emission = 668 nm)

Antibody Source

Polyclonal Rabbit IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all EGLN1/PHD2 Antibodies »

Product Specifications

Immunogen

The epitope recognized by this EGLN1/PHD2 antibody maps to a region between residues 1 and 50 of human PHD2/HIF Prolyl Hydroxylase 2 using the numbering given in entry NP_071334.1 (GeneID 54583).

Reactivity Notes

Results for use of this EGLN1/PHD2 antibody have been mixed in Rat with success in Western blot analysis and immunofluorescence on Rat endothelial cells and negative results with PC12 cells. Mouse reactivity reported in scientific literature (PMID: 25578858). Rat reactivity reported in scientific literature (PMID: 25635047). Primate reactivity reported in scientific literature (PMID: 25974097)

Localization

Cytoplasm, nucleus

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for EGLN1/PHD2 Antibody [Alexa Fluor® 647]

Flow Cytometry: EGLN1/PHD2 Antibody [Alexa Fluor® 647] [NB100-137AF647] - An intracellular stain was performed on Jurkat cells with EGLN1/PHD2 Antibody NB100-137AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Applications for EGLN1/PHD2 Antibody [Alexa Fluor® 647]

Application
Recommended Usage

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry

Optimal dilutions of this antibody should be experimentally determined.

Western Blot (Negative)

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
This PHD2 antibody is useful for Flow Cytometry, Immunocytochemistry/Immunofluorescence, Western Blot, and Immunohistochemistry-paraffin embedded sections. In ICC/IF, cytoplamic and nuclear staining was observed in HeLa cells. Immunoprecipitation was reported in scientific literature. 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.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

50mM Sodium Borate

Preservative

0.05% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C in the dark.

Background: EGLN1/PHD2

PHD2 (Prolyl Hydroxylase Domain-containing protein 2) belongs to the Prolyl-4-hydroxylase domain (PHD) family of proteins and is encoded by the Egl-9 Family Hypoxia Inducible Factor 1 (EGLN1) gene (1). Human EGLN1/PHD2 is a ubiquitously expressed enzyme that is 426 amino acids (aa) long with a theoretical molecular weight of ~46 kDa. Structurally PHD2 contains a nuclear export signal (NES, aa 6-20), an N-terminal MYND zinc finger domain (aa 21-58), and a C-terminal catalytic domain (aa 291-392) (2, 3). Functionally, PHD2 serves as an oxygen sensor and is responsible for post-translational modification of Hypoxia-inducible factor alpha (HIF-1alpha), a component of a transcriptional complex involved in oxygen homeostasis (1-3). During normoxia, PHD2 is responsible for oxygen-dependent hydroxylation of HIF-1alpha proline residue 402, 564, or both (3). The hydroxylation event promotes the binding of von Hippel-Lindau protein (VHL) and targets HIF1-alpha for ubiquitination and degradation (4, 5).

EGLN1/PHD2 has been implicated in several critical processes including erythropoiesis, angiogenesis, and metabolism as well as various pathologies such as cancer (2, 5, 6). Studies in mice have found that somatic deletion of PHD2 resulted in higher vascular endothelial growth factor A (VEGF-A) levels, increased blood vessel formation, and more erythropoietin (EPO), leading to severe polycythemia or erythrocytosis (high red blood cell (RBC) volume) (6). Another study revealed that specific point mutations in EGLN1/PHD2 led to elevated EPO and RBC mass associated with hemorrhages and strokes (6). Accordingly, given the known role of PHD2 in inhibition of EPO production, PHD2 inhibitors are being studied as a potential therapeutic for anemia (6). Additionally, dysregulation in EGLN1, and specifically the PHD2-VHL-HIF-1alpha pathway, has been associated with the development of pheochromocytomas (PCC) and sympathetic paragangliomas (PGL), which are rare neuroendocrine tumors (2). Besides pathological features, EGLN1/PHD2 may also be important for high altitude adaptation as two coding sequence variants in PHD2 are prevalent in the Tibetan population but is very rare in people at lower altitudes (2).

Alternate names for EGLN1/PHD2 include HIF Prolyl Hydroxylase 2, PH2, Prolyl hydroxylase domain containing protein 2, HIF2PH2, HIF-Prolyl hydroxylase 2, egl nine homolog 1, and C1orf12.

References

1. Amorim-Pires, D., Peixoto, J., & Lima, J. (2016). Hypoxia Pathway Mutations in Pheochromocytomas and Paragangliomas. Cytogenetic and genome research. https://doi.org/10.1159/000457479

2. Gardie, B., Percy, M. J., Hoogewijs, D., Chowdhury, R., Bento, C., Arsenault, P. R., Richard, S., Almeida, H., Ewing, J., Lambert, F., McMullin, M. F., Schofield, C. J., & Lee, F. S. (2014). The role of PHD2 mutations in the pathogenesis of erythrocytosis. Hypoxia (Auckland, N.Z.). https://doi.org/10.2147/HP.S54455

3. Minervini, G., Quaglia, F., & Tosatto, S. C. (2015). Insights into the proline hydroxylase (PHD) family, molecular evolution and its impact on human health. Biochimie. https://doi.org/10.1016/j.biochi.2015.07.009

4. Semenza G. L. (2007). Hypoxia-inducible factor 1 (HIF-1) pathway. Science's STKE : signal transduction knowledge environment. https://doi.org/10.1126/stke.4072007cm8

5. Chan, D. A., & Giaccia, A. J. (2010). PHD2 in tumour angiogenesis. British journal of cancer. https://doi.org/10.1038/sj.bjc.6605682

6. Meneses, A. M., & Wielockx, B. (2016). PHD2: from hypoxia regulation to disease progression. Hypoxia (Auckland, N.Z.). https://doi.org/10.2147/HP.S53576

Long Name

Egl Nine Homolog 1/Prolyl Hydroxylase Domain-containing Protein 2

Alternate Names

C1orf12, HIFPH2, HPH2, PHD2, SM20, ZMYND6

Gene Symbol

EGLN1

Additional EGLN1/PHD2 Products

Product Documents for EGLN1/PHD2 Antibody [Alexa Fluor® 647]

Product Datasheets

Certificate of Analysis

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

Download SDS

Product Specific Notices for EGLN1/PHD2 Antibody [Alexa Fluor® 647]

Alexa Fluor (R) products are provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment; (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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