製品の概要

  • 製品名Anti-PHD3 antibody
    PHD3 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to PHD3
  • アプリケーション適用あり: IP, ICC/IF, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human PHD3/prolyl hydroxylase.

    (Peptide available as ab30781.)

  • ポジティブ・コントロール
    • In WB this antibody gave a positive signal in whole cell, nuclear and cytoplasmic extracts of DFO-treated HeLa cells. In IF this antibody gave a positive signal in the following formaldehyde fixed cell line: HepG2.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab30782 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IP Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).

ターゲット情報

  • 機能Catalyzes the post-translational formation of 4-hydroxyproline in hypoxia-inducible factor (HIF) alpha proteins. Hydroxylates HIF-1 alpha at '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 in muscle cells and in apoptosis in neuronal tissue. Promotes cell death through a caspase-dependent mechanism.
  • 組織特異性Widely expressed at low levels. Expressed at higher levels in heart (cardiac myocytes, aortic endothelial cells and coronary artery smooth muscle) and placenta.
  • 配列類似性Contains 1 Fe2OG dioxygenase domain.
  • 細胞内局在Cytoplasm. Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Egl 9 family hypoxia inducible factor 3 antibody
    • Egl nine homolog 3 (C. elegans) antibody
    • Egl nine homolog 3 antibody
    • Egl nine like protein 3 isoform antibody
    • EGL9 homolog of C. elegans 3 antibody
    • EGLN3 antibody
    • EGLN3_HUMAN antibody
    • Factor responsive smooth muscle protein antibody
    • HIF Prolyl Hydroxylase 3 antibody
    • HIF-PH3 antibody
    • HIF-prolyl hydroxylase 3 antibody
    • HIFP4H3 antibody
    • HIFPH3 antibody
    • HPH-1 antibody
    • HPH-3 antibody
    • Hypoxia-inducible factor prolyl hydroxylase 3 antibody
    • P4H3 antibody
    • PHD3 antibody
    • Prolyl Hydroxylase Domain Containing Protein 3 antibody
    • Prolyl hydroxylase domain-containing protein 3 antibody
    • SM20 antibody
    see all

Anti-PHD3 antibody 画像

  • All lanes : Anti-PHD3 antibody (ab30782) at 1 µg/ml

    Lane 1 : Hela-Vehicle treated (Negative Control) Whole Cell Lysate (ab116321)
    Lane 2 : Hela-DFO treated (0.5mM, 24h) Whole Cell Lysate (ab116322)
    Lane 3 : Human HeLa Nuclear DFO treated
    Lane 4 : Human HeLa Cytoplasmic DFO treated

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 27 kDa
    Observed band size : 27 kDa
    Additional bands at : 18 kDa (possible non-specific binding).

    Exposure time : 20 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab30782 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

  • All lanes : Anti-PHD3 antibody (ab30782) at 1/1000 dilution

    Lane 1 : Mouse embroyonic fibroblast cell lysate. Treated 21% O2 for 24 hours
    Lane 2 : Mouse embroyonic fibroblast cell lysate. Treated 1% O2 for 24 hours

    Lysates/proteins at 5 µg per lane.

    Secondary
    Goat polyclonal anti-rabbit HRP-conjugate at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 27 kDa


    Exposure time : 30 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

  • ab30782 immunoprecipitating PHD3 from rat PC12 (whole cell lysate - the cells were either normoxia (21% oxygen) or hypoxia (1% hypoxia). A protein A matrix was used with the antibody at a dilution of 1/1000. A non-Abcam antibody was used in the subsequent western blot. The doublet observed is specific to PHD3 - most probably a modified form or an alternately spliced form. Note: This antibody IP's a hypoxia inducible species of ~27 kDa and ~30 kDa. This is the same molecular weight as the human and mouse isoform, not the SM-20 variant with the N-terminal mitochondrial leader sequence.

    Lane 1: normoxia - input

    Lane 2: hypoxia - input

    Lane 3: normoxia - control antibody IP

    Lane 4: hypoxia - control antibody IP

    Lane 5: normoxia - ab30782 IP

    Lane 6: hypoxia - ab30782 IP

    See Abreview

  • ICC/IF image of ab30782 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab30782 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Anti-PHD3 antibody (ab30782) 使用論文

This product has been referenced in:
  • Soler Palacios B  et al. Macrophages from the synovium of active rheumatoid arthritis exhibit an activin A-dependent pro-inflammatory profile. J Pathol 235:515-26 (2015). Read more (PubMed: 25319955) »
  • Van Welden S  et al. Differential expression of prolyl hydroxylase 1 in patients with ulcerative colitis versus patients with Crohn's disease/infectious colitis and healthy controls. J Inflamm (Lond) 10:36 (2013). WB, IHC-P ; Human . Read more (PubMed: 24257430) »

See all 6 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (mouse embryonic fibroblast)
Loading amount 5 µg
Specification mouse embryonic fibroblast
Treatment 21% O2 and 1% O2 for 24hrs
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Username

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投稿 May 17 2013

Thank you for contacting us.

As always, if we do not advise a certain method on our datasheet, the customer can use the one which they usually use within their lab. This is the case of ab110025.

For ab65979 I would suggest checking th...

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Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hearthe customer has had difficulty obtaining satisfactory results from this antibody.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Rat Cell lysate - whole cell (PC12)
Total protein in input 500 µg
Specification PC12
Treatment Normoxia and Hypoxia (1% Oxygen, 4 hrs)
Immuno-precipitation step Protein A
Username

Abcam user community

Verified customer

投稿 Oct 30 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (PC12)
Loading amount 40 µg
Specification PC12
Treatment Normoxia and Hypoxia (1% Oxygen, 4 hrs)
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Jun 20 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"