PHD1/prolyl hydroxylase Recombinant Rabbit mAb (S-5060-43)

Catalog Number: S0B60539 Application: WB, IHC-P Reactivity: Hu, Ms, Rt, Mk Conjugation: Unconjugated Brand: Starter

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$100 USD

Size: 25μl 100μl

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Product Details

Product Specification

Host	Rabbit
Antigen	PHD1/prolyl hydroxylase
Immunogen	Synthetic Peptide
Location	Nucleus
Accession	Q96KS0
Clone Number	S-5060-43
Antibody Type	Recombinant mAb
Isotype	IgG
Application	WB, IHC-P
Reactivity	Hu, Ms, Rt, Mk
Positive Sample	Ramos, Caco-2, A549, MCF7, HeLa, mouse brain, rat brain, rat heart, COS-7
Purification	Protein A
Concentration	2 mg/ml
Conjugation	Unconjugated
Physical Appearance	Liquid
Storage Buffer	PBS, 40% Glycerol, 0.05% BSA, 0.02% sodium azide
Stability & Storage	12 months from date of receipt / reconstitution, -20 °C as supplied

Dilution

application	dilution	species
WB	1:1000	Hu, Ms, Rt, Mk
IHC-P	1:200	Hu

Background

PHD1 (Prolyl Hydroxylase Domain-containing Protein 1), also known as EGLN2 or HIF-PH1, is a 2-oxoglutarate- and Fe²⁺-dependent prolyl hydroxylase that serves as a core component of the cellular oxygen sensor, catalyzing the hydroxylation of specific proline residues on hypoxia-inducible factor (HIF) α subunits under normoxic conditions, thereby mediating HIF degradation by the proteasome via the von Hippel-Lindau (VHL) ubiquitination complex. PHD1 prefers the C-terminal oxygen-dependent degradation domain (CODD) site for both HIF1A and HIF2A, and its activity is directly regulated by oxygen concentration, iron chelators, and cobalt ions, consistent with the functional characteristics of an oxygen sensor. Beyond HIF, PHD1 also hydroxylates CEP192, a key centrosomal component, linking its oxygen sensing to the cell cycle and primary cilia formation; it can also hydroxylate IKBKB, mediating NF-κB activation under hypoxic conditions. Under hypoxia, PHD1 hydroxylation activity is attenuated, allowing HIF to escape degradation and translocate into the nucleus, activating hypoxia-inducible gene expression; moreover, the PHD1 protein itself can be targeted for degradation under hypoxia by the SIAH1/SIAH2 ubiquitin ligases, constituting an additional layer of oxygen-sensing regulation.

Picture

Western Blot

WB result of PHD1/prolyl hydroxylase Recombinant Rabbit mAb
Primary antibody: PHD1/prolyl hydroxylase Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: Ramos whole cell lysate 20 µg
Lane 2: Caco-2 whole cell lysate 20 µg
Lane 3: A549 whole cell lysate 20 µg
Lane 4: MCF7 whole cell lysate 20 µg
Lane 5: HeLa whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 43 kDa
Observed MW: 48 kDa
This blot was developed with high sensitivity substrate

WB result of PHD1/prolyl hydroxylase Recombinant Rabbit mAb
Primary antibody: PHD1/prolyl hydroxylase Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: mouse brain lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 43 kDa
Observed MW: 43 kDa
This blot was developed with high sensitivity substrate

WB result of PHD1/prolyl hydroxylase Recombinant Rabbit mAb
Primary antibody: PHD1/prolyl hydroxylase Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: rat brain lysate 20 µg
Lane 2: rat heart lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 43 kDa
Observed MW: 48 kDa
This blot was developed with high sensitivity substrate

WB result of PHD1/prolyl hydroxylase Recombinant Rabbit mAb
Primary antibody: PHD1/prolyl hydroxylase Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: COS-7 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 43 kDa
Observed MW: 50 kDa
This blot was developed with high sensitivity substrate

Immunohistochemistry

IHC shows positive staining in paraffin-embedded human testis. Anti-PHD1 antibody was used at 1/200 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human endometrial cancer. Anti-PHD1 antibody was used at 1/200 dilution, followed by a HRP Polymer for Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.