使用敲除细胞株进行验证RabMAb

Anti-PARP抗体[E102] (ab32138)

概述

  • 产品名称Anti-PARP抗体[E102]
    参阅全部 PARP 一抗
  • 描述
    兔单克隆抗体[E102] to PARP
  • 特异性ab32138 recognises both pro-form and p25 cleaved form of PARP.
  • 经测试应用适用于: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human PARP (N terminal). The exact sequence is proprietary.

  • 阳性对照
    • Jurkat cell lysate, human brain tissue
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

性能

应用

Our Abpromise guarantee covers the use of ab32138 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 1/1000 - 1/10000. Predicted molecular weight: 113 kDa.

Existing as a 113 kDa nuclear protein, PARP is cleaved between amino acids Asp214 and Gly215 to yield two fragments of 29 kDa (C-terminal catalytic domain) and 85 kDa (N-terminal DNA-binding domain)

IHC-P 1/25.
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100 - 1/200.

Use with paraformaldehyde fixed cells.

靶标

  • 功能Involved in the base excision repair (BER) pathway, by catalyzing the poly(ADP-ribosyl)ation of a limited number of acceptor proteins involved in chromatin architecture and in DNA metabolism. This modification follows DNA damages and appears as an obligatory step in a detection/signaling pathway leading to the reparation of DNA strand breaks. Mediates the poly(ADP-ribosyl)ation of APLF and CHFR. Positively regulates the transcription of MTUS1 and negatively regulates the transcription of MTUS2/TIP150.
  • 序列相似性Contains 1 BRCT domain.
    Contains 1 PARP alpha-helical domain.
    Contains 1 PARP catalytic domain.
    Contains 2 PARP-type zinc fingers.
  • 翻译后修饰Phosphorylated by PRKDC. Phosphorylated upon DNA damage, probably by ATM or ATR.
    Poly-ADP-ribosylated by PARP2. Poly-ADP-ribosylation mediates the recruitment of CHD1L to DNA damage sites.
    S-nitrosylated, leading to inhibit transcription regulation activity.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • ADP ribosyltransferase (NAD+; poly (ADP ribose) polymerase) antibody
    • ADP ribosyltransferase diphtheria toxin like 1 antibody
    • ADP ribosyltransferase NAD(+) antibody
    • ADPRT 1 antibody
    • ADPRT antibody
    • ADPRT1 antibody
    • ARTD1 antibody
    • msPARP antibody
    • NAD(+) ADP ribosyltransferase 1 antibody
    • NAD(+) ADP-ribosyltransferase 1 antibody
    • pADPRT 1 antibody
    • pADPRT1 antibody
    • PARP 1 antibody
    • PARP antibody
    • PARP-1 antibody
    • PARP1 antibody
    • PARP1_HUMAN antibody
    • Poly (ADP ribose) polymerase 1 antibody
    • poly (ADP ribose) polymerase family, member 1 antibody
    • Poly [ADP-ribose] polymerase 1 antibody
    • Poly(ADP ribose) polymerase antibody
    • poly(ADP ribose) synthetase antibody
    • poly(ADP ribosyl)transferase antibody
    • Poly[ADP ribose] synthetase 1 antibody
    • Poly[ADP-ribose] synthase 1 antibody
    • PPOL antibody
    see all

Anti-PARP antibody [E102] 图像



  • Predicted band size : 113 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: PARP1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: MCF7 whole cell lysate (20 µg)                                                               

    Lanes 1 - 4: Merged signal (red and green). Green - ab32138 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.                                                   

    ab32138 was shown to specifically react with PARP1 when PARP1 knockout samples were used. Wild-type and PARP1 knockout samples were subjected to SDS-PAGE.  Ab32138 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10 000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10 000 dilution for 1 hour at room temperature before imaging.

  • ab32138 (1/200) staining PARP in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised in 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.

    See Abreview

  • All lanes : Anti-PARP antibody [E102] (ab32138) at 1/1000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : Jurkat + Camptothecin cell lysate


    Predicted band size : 113 kDa
    Observed band size : 25,120 kDa (why is the actual band size different from the predicted?)
  • Immunohistochemical analysis of PARP expression in paraffin embedded human brain tissue section, using 1/25 ab32138.

  • Overlay histogram showing Jurkat cells stained with ab32138 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32138, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1:500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Anti-PARP antibody [E102] (ab32138)参考文献

This product has been referenced in:
  • Shen C  et al. BRCA1-associated protein 1 deficiency in lung adenocarcinoma predicts poor outcome and increased tumor invasion. BMC Cancer 16:670 (2016). WB . Read more (PubMed: 27553041) »
  • Cao C  et al. The long intergenic noncoding RNA UFC1, a target of MicroRNA 34a, interacts with the mRNA stabilizing protein HuR to increase levels of ß-catenin in HCC cells. Gastroenterology 148:415-26.e18 (2015). WB . Read more (PubMed: 25449213) »

See all 7 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 Tissue lysate - whole (Kidney)
Loading amount 56 µg
Specification Kidney
Gel Running Conditions Reduced Denaturing (4-20% Tris Glycin)
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 May 16 2013

Thank you for your inquiry.
The concentration of lot GR29754-10 is 0.097 mg/ml.
I hope this information helps. Please contact us with any other questions.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton-X100 in PBS
Username

Dr. Kirk McManus

Verified customer

提交于 May 31 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Intestine)
Specification Intestine
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Username

Abcam user community

Verified customer

提交于 May 16 2012

We loaded 15 ug of protein from Jurkat onto Western blot. I hope this is helpful. Please contact us again if you have any further questions.

Thank you for your telephone enquiry yesterday. I have done a literature search to investigate the different cleavage products of PARP. There is a reference in which analysis of bovine PARP reveals three sites with high homology to known MMP-2 clea...

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