Datasheet

生物学的記述

Specificity	PHD1 Antibody (Rabbit mAb) [F19A8] detects endogenous levels of total PHD1 protein.
Background	PHD1, also designated EGLN2, belongs to the family of 2-oxoglutarate- and iron-dependent dioxygenases that function collectively, alongside PHD2 and PHD3, as the core oxygen-sensing machinery of metazoan cells, hydroxylating specific proline residues within a conserved LXXLAP recognition motif found in target proteins. PHD1 hydroxylates HIF1A and HIF2A at conserved proline residues located in each subunit's N-terminal and C-terminal oxygen-dependent degradation domains, and only one of these two prolines needs to be hydroxylated to trigger destruction of the HIF-alpha subunit, with hydroxylation at the C-terminal site occurring preferentially before modification at the N-terminal site. This proline hydroxylation reaction requires molecular oxygen as a direct cosubstrate, making PHD1 catalytically active only when sufficient oxygen is present, and the hydroxylated proline creates the recognition surface engaged by the von Hippel-Lindau protein, the substrate-recognition component of an Elongin BC-Cullin 2-Rbx1 E3 ubiquitin ligase complex that ubiquitinates HIF-alpha and targets it for proteasomal degradation, so that under normal oxygen tension HIF-alpha is continuously synthesized and destroyed while hypoxia removes PHD1's oxygen cosubstrate and allows HIF-alpha to escape degradation and accumulate. Beyond its canonical role in HIF regulation, PHD1 hydroxylates a distinct set of non-HIF substrates, including ATF4, IKBKB, and CEP192, extending its proline-hydroxylase activity into transcriptional and cell-cycle regulatory pathways outside the oxygen-sensing axis; PHD1-mediated hydroxylation of the transcription factor FOXO3a specifically destabilizes FOXO3a by blocking its interaction with the deubiquitinase USP9X, providing one defined mechanistic route through which PHD1 activity influences cell survival signaling independently of HIF. Ectopic expression of PHD1 suppresses HIF-1alpha accumulation and reduces secretion of vascular endothelial growth factor following hypoxic stimulation, and tumors formed by colon carcinoma cells engineered to express PHD1 show inhibited growth, increased necrosis, and reduced microvessel density when implanted in mice, directly linking restored PHD1 hydroxylase activity to suppression of the angiogenic HIF output that tumors otherwise depend on under hypoxic stress.

Specificity

PHD1 Antibody (Rabbit mAb) [F19A8] detects endogenous levels of total PHD1 protein.

Background

PHD1, also designated EGLN2, belongs to the family of 2-oxoglutarate- and iron-dependent dioxygenases that function collectively, alongside PHD2 and PHD3, as the core oxygen-sensing machinery of metazoan cells, hydroxylating specific proline residues within a conserved LXXLAP recognition motif found in target proteins. PHD1 hydroxylates HIF1A and HIF2A at conserved proline residues located in each subunit's N-terminal and C-terminal oxygen-dependent degradation domains, and only one of these two prolines needs to be hydroxylated to trigger destruction of the HIF-alpha subunit, with hydroxylation at the C-terminal site occurring preferentially before modification at the N-terminal site. This proline hydroxylation reaction requires molecular oxygen as a direct cosubstrate, making PHD1 catalytically active only when sufficient oxygen is present, and the hydroxylated proline creates the recognition surface engaged by the von Hippel-Lindau protein, the substrate-recognition component of an Elongin BC-Cullin 2-Rbx1 E3 ubiquitin ligase complex that ubiquitinates HIF-alpha and targets it for proteasomal degradation, so that under normal oxygen tension HIF-alpha is continuously synthesized and destroyed while hypoxia removes PHD1's oxygen cosubstrate and allows HIF-alpha to escape degradation and accumulate. Beyond its canonical role in HIF regulation, PHD1 hydroxylates a distinct set of non-HIF substrates, including ATF4, IKBKB, and CEP192, extending its proline-hydroxylase activity into transcriptional and cell-cycle regulatory pathways outside the oxygen-sensing axis; PHD1-mediated hydroxylation of the transcription factor FOXO3a specifically destabilizes FOXO3a by blocking its interaction with the deubiquitinase USP9X, providing one defined mechanistic route through which PHD1 activity influences cell survival signaling independently of HIF. Ectopic expression of PHD1 suppresses HIF-1alpha accumulation and reduces secretion of vascular endothelial growth factor following hypoxic stimulation, and tumors formed by colon carcinoma cells engineered to express PHD1 show inhibited growth, increased necrosis, and reduced microvessel density when implanted in mice, directly linking restored PHD1 hydroxylase activity to suppression of the angiogenic HIF output that tumors otherwise depend on under hypoxic stress.

使用情報

Application

WB, IHC, IF, FCM

Dilution

WB	IHC	IF	FCM
1:1000-1:10000	1:100 - 1:250	1:100 - 1:250	1:10 - 1:100

Reactivity

Human

Source

Rabbit Monoclonal Antibody

MW

44 kDa

Storage Buffer

PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage
(from the date of receipt)

-20°C (avoid freeze-thaw cycles), 2 years

プロトコール

References

https://pubmed.ncbi.nlm.nih.gov/28622522/
https://pubmed.ncbi.nlm.nih.gov/14695194/

PHD1 Antibody (Rabbit mAb) [F19A8]

生物学的記述

使用情報

References

Application Data