Name: Anti-PHD1/prolyl hydroxylase antibody [EPR2746]
Brand: RabMAb
SKU: ab113077
Rating: 4 (2 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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

Carrier Free

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.

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

Alternative Versions
- Anti-PHD1/prolyl hydroxylase antibody [EPR2746] - BSA and Azide free (ab242386)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- HeLa nuclear extract lysate (ab14655)
Recombinant Protein
- Recombinant Human PHD1/prolyl hydroxylase protein (ab132272)

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.

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

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 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) 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) 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)

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)

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)

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)

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)

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)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

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

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

Abcam user community

Verified customer

Submitted Oct 20 2016

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

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!

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.

Abcam Scientific Support

Answered on Oct 26 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

References (2)

Customer reviews and Q&As

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

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