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  1. Link

    phd1prolyl-hydroxylase-antibody-epr2746-ab113077.pdf

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Cardiovascular Hypoxia Prolyl Hydroxylase
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RecombinantRabMAb

Recombinant Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)

  • Datasheet
  • SDS
Reviews (2)Q&A (1)References (2)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
  • Immunocytochemistry/ Immunofluorescence - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
  • Flow Cytometry (Intracellular) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
  • Western blot - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
  • Western blot - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
  • Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2746] to PHD1/prolyl hydroxylase
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-PHD1/prolyl hydroxylase antibody [EPR2746]
    See all PHD1/prolyl hydroxylase primary antibodies
  • Description

    Rabbit monoclonal [EPR2746] to PHD1/prolyl hydroxylase
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A549, HeLa and MCF-7 cell lysates, Human fetal heart, Mouse brain, Mouse heart, Rat brain, Rat heart tissue lysates IHC-P: Mouse testis, Rat spleen, Human breast carcinoma and Human lung carcinoma tissue ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR2746
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Hypoxia
    • Prolyl Hydroxylase
    • Epigenetics and Nuclear Signaling
    • Cardiovascular/Immune
    • Hypoxia
    • Prolyl hydroxylase
    • Cancer
    • Invasion/microenvironment
    • Hypoxia
    • Hydroxylases
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Anti-PHD1/prolyl hydroxylase antibody [EPR2746] - BSA and Azide free (ab242386)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human PHD1/prolyl hydroxylase protein (ab132272)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab113077 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt (Intra)
1/30.
WB (2)
1/1000 - 1/10000. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

ICC/IF
1/50 - 1/100.
Notes
Flow Cyt (Intra)
1/30.
WB
1/1000 - 1/10000. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.

ICC/IF
1/50 - 1/100.

Target

  • Function

    Catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates HIF-1 alpha at 'Pro-402' and 'Pro-564', and HIF-2 alpha. Functions as a cellular oxygen sensor and, under normoxic conditions, targets HIF through the hydroxylation for proteasomal degradation via the von Hippel-Lindau ubiquitination complex. May play a role in cell growth regulation. Isoform p40 and isoform p43 exhibit the same level of activity.
  • Tissue specificity

    Expressed abundantly in all tissues with highest expression in testis. Expressed in hormone responsive tissues, including normal and cancerous mammary, ovarian and prostate epithelium.
  • Sequence similarities

    Contains 1 Fe2OG dioxygenase domain.
  • Cellular localization

    Cytoplasm. Nucleus.
  • Target information above from: UniProt accession Q96KS0 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 112398 Human
    • Entrez Gene: 112406 Mouse
    • Entrez Gene: 308457 Rat
    • Omim: 606424 Human
    • SwissProt: Q96KS0 Human
    • SwissProt: Q91YE2 Mouse
    • SwissProt: Q6AYU4 Rat
    • Unigene: 515417 Human
    • Unigene: 29978 Mouse
    see all
  • Alternative names

    • 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
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat spleen tissue sections labeling PHD1/prolyl hydroxylase with Purified ab113077 at 1:50 dilution (5.14 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    Immunocytochemistry/ Immunofluorescence - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling PHD1/prolyl hydroxylase with purified ab113077 at 1/50 dilution (5 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL) was used as the secondary antibody only control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse testis tissue sections labeling PHD1/prolyl hydroxylase with Purified ab113077 at 1:50 dilution (5.14 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling PHD1/prolyl hydroxylase with Purified ab113077 at 1:50 dilution (5.14 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Flow Cytometry (Intracellular) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    Flow Cytometry (Intracellular) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)

    Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PHD1/prolyl hydroxylase with Purified ab113077 at 1/30 dilution (10µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 

     

     

  • Western blot - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    Western blot - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    All lanes : Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077) at 1/1000 dilution (Purified)

    Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Human fetal heart lysates
    Lane 3 : Mouse brain lysates
    Lane 4 : Mouse heart lysates
    Lane 5 : Rat brain lysates
    Lane 6 : Rat heart lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 44 kDa
    Observed band size: 40,43 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    Western blot - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    All lanes : Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077) at 1/1000 dilution

    Lane 1 : A549 cell lysates
    Lane 2 : HeLa cell lysates
    Lane 3 : MCF-7 cell lysates

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 44 kDa



    This image was generated using the unpurified version of the product. 

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)

    ab113077, at a 1/100 dilution, staining PHD1/prolyl hydroxylase in paraffin-embedded Human breast carcinoma tissue by Immunohistochemistry. 

    This image was generated using the unpurified version of the product. 

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)

    ab113077, at a 1/100 dilution, staining PHD1/prolyl hydroxylase in paraffin-embedded Human lung carcinoma tissue by Immunohistochemistry.

     This image was generated using the unpurified version of the product. 

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)
    Anti-PHD1/prolyl hydroxylase antibody [EPR2746] (ab113077)

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (2)

Publishing research using ab113077? Please let us know so that we can cite the reference in this datasheet.

ab113077 has been referenced in 2 publications.

  • Tsai YM  et al. Loss of miR-145-5p Causes Ceruloplasmin Interference with PHD-Iron Axis and HIF-2a Stabilization in Lung Adenocarcinoma-Mediated Angiogenesis. Int J Mol Sci 21:N/A (2020). PubMed: 32708433
  • Zurlo G  et al. Prolyl hydroxylase substrate adenylosuccinate lyase is an oncogenic driver in triple negative breast cancer. Nat Commun 10:5177 (2019). PubMed: 31729379

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Western blot abreview for Anti-PHD1/prolyl hydroxylase antibody [EPR2746]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (MKN7, MKN45, SNU-1 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
MKN7, MKN45, SNU-1 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 20 2016

Western blot abreview for Anti-PHD1/prolyl hydroxylase antibody [EPR2746]

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - whole cell (osteosarcoma cells)
Specification
osteosarcoma cells
Treatment
control and siRNA for PHD1
Blocking step
Milk as blocking agent for 15 minute(s) · Concentration: 10% · Temperature: 21°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Sonia Rocha

Verified customer

Submitted Feb 13 2015

Question

Dear Sir/Madam:
Recently, I ordered two different anti-PHD rabbit monoclonal
antibodies (ab109088 and ab113077). The datasheet states that they
have to be stored at -20°C. According to the documents, the antibodies
were shipped on 19/10. They were received in this lab on monday 22/10.
We accidentally stored them at 4°C until today. Is this an important
problem or will the antibodies still work?
Thank you for the information!

Read More

Abcam community

Verified customer

Asked on Oct 26 2012

Answer

Thank you for contacting us. I would like to reassure you the quality of these antibodies may not have been affected by remaining at 4C during a week.

We recently undertook an in-depth study examining storage conditions and antibody performance. After over 13,000 individual ELISA experiments, we have determined that storing our antibodies at various temperatures of up to 45°C for 1 week does not impact their activity. I would be happy to share a summary of our findings with you; please find the results attached.

It is important however making sure the antibody is now store at – 20C as well as avoiding freeze / thaw cycles.


I hope this information helps, but please don't hesitate to contact me for additional information or assistance.

Read More

Abcam Scientific Support

Answered on Oct 26 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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