PRODUCT CODE: ET1604-5

PHD1/prolyl hydroxylase Recombinant Rabbit Monoclonal Antibody [SP00-48] (ET1604-5)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of PHD1/prolyl hydroxylase on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1604-5, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Hela cell lysate<br />
Lane 2: PC-12 cell lysate<br />
Lane 2: NIH/3T3 cell lysate
  • Western blot analysis of PHD1/prolyl hydroxylase on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1604-5, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Hela cell lysate<br />
Lane 2: PC-12 cell lysate<br />
Lane 2: NIH/3T3 cell lysate
  • ICC staining of PHD1/prolyl hydroxylase in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1604-5, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of PHD1/prolyl hydroxylase in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1604-5, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of PHD1/prolyl hydroxylase in SKOV-3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1604-5, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-PHD1/prolyl hydroxylase antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-5, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-PHD1/prolyl hydroxylase antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-5, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-PHD1/prolyl hydroxylase antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-5, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse prostate tissue using anti-PHD1/prolyl hydroxylase antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1604-5, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of PHD1/prolyl hydroxylase was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1604-5, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
Western blot analysis of PHD1/prolyl hydroxylase on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1604-5, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: PC-12 cell lysate
Lane 2: NIH/3T3 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

PHD1/prolyl hydroxylase Recombinant Rabbit Monoclonal Antibody [SP00-48] (ET1604-5)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

A549, SKOV-3, Hela, PC12, NIH/3T3, mouse prostate tissue, human lung cancer tissue, human breast carcinoma tissue, mouse testis tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SP00-48

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

44 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:5,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

PHD1/prolyl hydroxylase

SYNONYMS

DKFZp434E026 antibody; EGL nine (C.elegans) homolog 2 antibody; Egl nine homolog 2 (C. elegans) antibody; Egl nine homolog 2 antibody; EGLN 2 antibody; EGLN2 antibody; EGLN2_HUMAN antibody; EIT 6 antibody; EIT6 antibody; Estrogen-induced tag 6 antibody; HIF P4H 1 antibody; HIF PH1 antibody; HIF prolyl hydroxylase 1 antibody; HIF-PH1 antibody; HIF-prolyl hydroxylase 1 antibody; HIFPH 1 antibody; HIFPH1 antibody; HPH 3 antibody; HPH-1 antibody; HPH-3 antibody; HPH3 antibody; Hypoxia inducible factor prolyl hydroxylase 1 antibody; Hypoxia-inducible factor prolyl hydroxylase 1 antibody; P4H1 antibody; PHD 1 antibody; PhD1 antibody; prolyl hydroxylase domain containing protein 1 antibody; Prolyl hydroxylase domain-containing protein 1 antibody

TISSUE SPECIFICITY

Expressed in adult and fetal heart, brain, liver, lung, skeletal muscle, and kidney. Also expressed in testis and placenta. Highest levels in adult brain, placenta, lung, kidney, and testis. Expressed in hormone responsive tissues, including normal and cancerous mammary, ovarian and prostate epithelium.

POST-TRANSLATIONAL MODIFICATION

Ubiquitinated by SIAH1 and/or SIAH2 in response to the unfolded protein response (UPR), leading to its degradation.

SUBCELLULAR LOCATION

Nucleus, cytosol.

FUNCTION

Prolyl hydroxylase domain proteins HIF PHD1, HIF PHD2 and HIF PHD3 (known as PHD1, PHD2 and PHD3 in rodents, respectively) can hydroxylate HIF-α subunits. Hypoxia-inducible factor (HIF) is a transcriptional regulator important in several aspects of oxygen homeostasis. The prolyl hydroxylases catalyze the posttranslational formation of 4-hydroxyproline in HIF-α proteins. HIF PHD1, which is widely expressed, with highest levels of expression in testis, functions as a cellular oxygen sensor and is important in cell growth regulation. HIF PHD1 can localize to the nucleus or the cytoplasm and is also detected in hormone responsive tissues, such as normal and cancerous mammary, ovarian and prostate epithelium. HIF PHD1 is encoded by EGLN2, which maps to chromosome 19q13.3. HIF PHD2 is regarded as the main cellular oxygen sensor, as RNA interference against HIF PHD2, but not HIF PHD1 or HIF PHD3, is enough to stabilize HIF-1α in normoxia. HIF PHD2, a direct HIF target gene, is expressed mainly in skeletal muscle, heart, kidney and brain. HIF PHD3 may play a role in the regulation of cell growth in muscle cells and in apoptosis in neuronal tissue. HIF PHD3 is widely expressed, although the highest levels can be detected in placenta and he