Anti-Poly (ADP-Ribose) Polymer antibody [10H] (ab14459)

Overview

  • Product name
    Anti-Poly (ADP-Ribose) Polymer antibody [10H]
    See all Poly (ADP-Ribose) Polymer primary antibodies
  • Description
    Mouse monoclonal [10H] to Poly (ADP-Ribose) Polymer
  • Specificity
    This antibody reacts with poly (ADP-Ribose) Polymer synthesized by a variety of poly(ADP- ribose) polymerases (PARP)-related enzymes including PARP1, 2, 3, tankyrase, vPARP, sPARP and others. The antibody does not cross-react with ADP-ribose, 5'-AMP, or yeast RNA as tested by ELISA.
  • Tested applications
    Suitable for: WB, ELISA, ICC/IF, IHC-Fr, Flow Cytmore details
  • Immunogen

    Other Immunogen Type. Poly (ADP-Ribose) polymer mixed with methylated bovine serum albumin.

  • Positive control
    • Rat liver induced for Poly (ADP-Ribose) Polymer synthesis by injection with diethylnitrosamine. MEF's treated with 500um Hydrogen Peroxide.

Properties

Applications

Our Abpromise guarantee covers the use of ab14459 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
WB Use a concentration of 2 - 10 µg/ml. 2 µg/ml, if using ECL or 10 µg/ml, if using colorimetric methods.
ELISA Use at an assay dependent concentration.
ICC/IF 1/400.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab18392 - Mouse monoclonal IgG3, is suitable for use as an isotype control with this antibody.

Target

  • Relevance
    Poly (ADP-Ribose) is a polymer synthesized by a class of enzymes named poly(ADP-ribose) polymerases (PARP). Using NAD+ as substrate, PARP catalyzes the formation of the polymer poly (ADP-Ribose), with chain lengths ranging from 2 to 300 residues, containing approximately 2% branching in the chain. Poly (ADP-Ribose) polymer becomes attached to nuclear proteins, and to PARP itself (automodification). Under normal conditions, cells display low basal level of poly (ADP-Ribose) polymer, which can dramatically increase in cells exposed to DNA damaging agents (irradiation, alkylation, etc.). This increase of polymer synthesis is usually transient and is followed by a rapid degradation phase with a short half life which can be less than 1 min. The low endogenous level of polymer in unstimulated cells and its rapid catabolism during DNA damage has been ascribed to high activity of the polymer catabolizing enzyme poly(ADP-ribose) glycohydrolyase (PARG).
  • Alternative names
    • pADPr antibody
    • Poly ADP ribose antibody

Images

  • All lanes : Anti-Poly (ADP-Ribose) Polymer antibody [10H] (ab14459) at 1/1000 dilution

    Lane 1 : Ladder
    Lane 2 : Ladder
    Lane 3 : Whole cell lysate prepared from MEF's
    Lane 4 : Whole cell lysate prepared from MEF's treated with 500um Hydrogen Peroxide, 10 minutes
    Lane 5 : Whole cell lysate prepared from MEF's treated with 500um Hydrogen Peroxide, 20 minutes
    Lane 6 : Whole cell lysate prepared from MEF's treated with 500um Hydrogen Peroxide, 30 minutes

    Secondary
    IRDye 800CW conjugated goat monoclonal at 1/10000 dilution

    Image courtesy of Dr Aashish Joshi by Abreview.

    See Abreview

  • Immunohistochemistry of rat livers treated with diethylnitrosamine (200 mg/kg) and stained with ab14459 diluted 1/100. After treatment livers were removed and rapidly processed 10 hr later, at peak polymer induction. Left hand side image was from diethylnitrosamine untreated liver tissue and right one represents DEN treated sections.
  • ab14459 staining Poly (ADP-Ribose) Polymer in Rat cardiomyoblast (H9c2) by Flow Cytometry. Cells were fixed with formaldehyde and permeabilized with 0.1% Triton X-100. The sample was incubated with the primary antibody (1/1000 in 3% BSA) for 1 hour at 37°C. ab98707, FITC-conjugated Goat anti-mouse FITC (IgG3 heavy chain preadsorbed 1/1000) was used as the secondary antibody.

    See Abreview

  • ab14459 staining Poly (ADP-Ribose) Polymer in human breast cancer cells (MCF7 cells) by Immunocytochemistry/ Immunofluorescence. The cells were paraformaldehyde fixed, permeabilised in 0.2% Triton/ PBS and then blocked using 1% BSA and 3% FBS in PBS for 30 minutes at room temperature. Samples were then incubated with primary antibody at 1/50 for 1 hour. The secondary antibody used was a goat anti-mouse IgG conjugated to Alexa Fluor® 555 (red) used at a 1/500 dilution.

    See Abreview

References

This product has been referenced in:
  • Xiang C  et al. LncRNA-SLC6A9-5:2: A potent sensitizer in 131I-resistant papillary thyroid carcinoma with PARP-1 induction. Oncotarget 8:22954-22967 (2017). WB ; Human . Read more (PubMed: 28086241) »
  • Eros K  et al. Chronic PARP-1 inhibition reduces carotid vessel remodeling and oxidative damage of the dorsal hippocampus in spontaneously hypertensive rats. PLoS One 12:e0174401 (2017). Read more (PubMed: 28339485) »

See all 16 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
20 µg
Specification
U2OS cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
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Submitted Sep 18 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (human tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM NaCitrate ph6
Permeabilization
No
Specification
human tonsil
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 27°C
Fixative
Formaldehyde
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Submitted Mar 06 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (GONAD)
Antigen retrieval step
Enzymatic
Permeabilization
No
Specification
GONAD
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative
Paraformaldehyde
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Submitted Feb 10 2014

Application
Western blot
Sample
Human Cell lysate - whole cell (vascular smooth muscle cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
UV irradiation 20 J/m2 (30 mins recovery)
Specification
vascular smooth muscle cell
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted Feb 06 2014

Application
Western blot
Sample
Human Cell lysate - whole cell (Breast Cancer cells)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
30 µg
Specification
Breast Cancer cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Nov 04 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Rat Cell (Cardiomyoblast (H9c2))
Permeabilization
Yes - Triton 100x 0,1%
Gating Strategy
All
Specification
Cardiomyoblast (H9c2)
Preparation
Cell harvesting/tissue preparation method: standart trypsinization
Sample buffer: PBS
Fixation
Formaldehyde
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Submitted Sep 16 2013

Thank you for your message. I am very pleased to hear you would like to accept our offer and test ab14459 and ab14460 in IP.

ab14459
DISCOUNT CODE: ######
Expiration date: 4th Feb 2013

ab14460
DISCOUNT CODE: #######...

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Thank you for your message. i hope you enjoyed the EMBO conference.

I am sorry to confirm that to our knowledge, ab14459 and ab14460 have not been tested in IP. They have been tested in western blotting.

Therefore, I can offer a d...

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I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab14459.

To check the status of the order please contact o...

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I would be happy to offer a replacement antibody for the vial of ab14459 that is not working for you.

Please provide me with the PO number or Abcam order reference number associated with the purchase of the faulty product, as well as the cata...

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