概述

  • 产品名称Anti-PADPR抗体[10H]
    参阅全部 PADPR 一抗
  • 描述
    小鼠单克隆抗体[10H] to PADPR
  • 特异性This antibody reacts with PADPR 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.
  • 经测试应用适用于: WB, ELISA, ICC/IF, IHC-Fr, Flow Cytmore details
  • 种属反应性
    与反应: Rat, Human
  • 免疫原

    PADPR mixed with methylated bovine serum albumin.

  • 阳性对照
    • Rat liver induced for PADPR synthesis by injection with diethylnitrosamine. MEF's treated with 500um Hydrogen Peroxide.

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 相关性PADPR (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 PADPR, with chain lengths ranging from 2 to 300 residues, containing approximately 2% branching in the chain. PADPR becomes attached to nuclear proteins, and to PARP itself (automodification). Under normal conditions, cells display low basal level of PADPR 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).
  • 别名
    • Poly ADP ribose antibody

Anti-PADPR antibody [10H] 图像

  • ab14459 staining PADPR 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

  • 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 PADPR 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

  • All lanes : Anti-PADPR 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

Anti-PADPR antibody [10H] (ab14459)参考文献

This product has been referenced in:
  • Li X  et al. Histone demethylase KDM5B is a key regulator of genome stability. Proc Natl Acad Sci U S A 111:7096-101 (2014). Read more (PubMed: 24778210) »
  • Guillot C  et al. PARP inhibition and the radiosensitizing effects of the PARP inhibitor ABT-888 in in vitro hepatocellular carcinoma models. BMC Cancer 14:603 (2014). WB ; Human . Read more (PubMed: 25139788) »

See all 7 Publications for this product

Product Wall

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

Abcam user community

Verified customer

提交于 Feb 10 2014

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

Abcam user community

Verified customer

提交于 Feb 06 2014

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

Abcam user community

Verified customer

提交于 Nov 04 2013

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

Abcam user community

Verified customer

提交于 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: #######...

Read More

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...

Read More

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...

Read More

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...

Read More
Application Western blot
Sample Mouse Cell lysate - whole cell (MEFs)
Loading amount 100 µg
Specification MEFs
Treatment Hydrogen Peroxide 500uM
Gel Running Conditions Reduced Denaturing (Gradient gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Dr. AASHISH JOSHI

Verified customer

提交于 Feb 18 2012

Thanks for your reply.

That information was helpful. But I'm still not quite sure what the problem is. It seems you are doing western blotting. Are you seeing a band at an unexpected molecular weight? Are you seeing high background? No bands?...

Read More

1-10 of 18 Abreviews or Q&A

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