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
  • Host species
    Mouse
  • 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 conjugated to bovine serum albumin. 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

    Lanes 1-2 : 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
    All lanes : IRDye 800CW conjugated goat monoclonal at 1/10000 dilution

    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:
  • Mao Y  et al. PARP inhibitor olaparib sensitizes cholangiocarcinoma cells to radiation. Cancer Med 7:1285-1296 (2018). WB ; Human . Read more (PubMed: 29479816) »
  • Fliegert R  et al. 2'-Deoxyadenosine 5'-diphosphoribose is an endogenous TRPM2 superagonist. Nat Chem Biol 13:1036-1044 (2017). IP ; Human . Read more (PubMed: 28671679) »
See all 19 Publications for this product

Customer reviews and Q&As

1-10 of 21 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Feb 05 2018

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

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

Abcam user community

Verified customer

Submitted Sep 16 2013

Answer

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: #######
Expiration date: 4th Feb 2013

Each code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ab14459 and ab14460 in IP and include the corresponding code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.

For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreviews and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More

Answer

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 discount off a future purchase if you buy ab14459 and ab14460 now, testthem in IP and submit feedback to us in the form of an Abreview. It doesn't matter whether the Abreview is positive or negative, we would just really like to receive your feedback. You would recieve one discount code for each antibody. Eachdiscount codewould be to the value of: 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab14459 and ab14460 in IP. I will then send a discount code. This code must be issued before purchasingthe antibodiesso please wait for my reply before ordering.

2. Purchase ab14459 and ab14460 either by phone, fax, or online (https://www.abcam.com). This purchase will be at full price.

3. Test the antibodiesin IP.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code.Each discount codecan be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.

Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab14459or ab14460 turns out to be unsuitable for IP, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More

Question
Answer

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 our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

1-10 of 21 Abreviews or Q&A

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

Sign up