Overview

  • Product name
    Anti-PARP1 antibody [E102]
    See all PARP1 primary antibodies
  • Description
    Rabbit monoclonal [E102] to PARP1
  • Host species
    Rabbit
  • Specificity
    ab32138 recognises both pro-form and p25 cleaved form of PARP1.
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human PARP1 (N terminal). The exact sequence is proprietary.

  • Positive control
    • WB: Wild type HAP1 whole cell lysate; HeLa whole cell lysate; Jurkat cell lysate. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt: Jurkat cells.
  • General notes

    A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.

Application Abreviews Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 113 kDa.

Existing as a 113 kDa nuclear protein, PARP1 is cleaved between amino acids Asp214 and Gly215 to yield two fragments of 29 kDa (N-terminal catalytic domain) and 85 kDa (C-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.

Target

  • Function
    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.
  • Sequence similarities
    Contains 1 BRCT domain.
    Contains 1 PARP alpha-helical domain.
    Contains 1 PARP catalytic domain.
    Contains 2 PARP-type zinc fingers.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • ADP ribosyltransferase (NAD+; poly (ADP ribose) polymerase) antibody
    • ADP ribosyltransferase 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

Images

  • 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.

  • Immunohistochemical analysis of PARP1 expression in paraffin embedded human brain tissue section, using 1/25 ab32138.

  • ab32138 (1/200) staining PARP1 in HeLa (human epithelial cell line from cervix adenocarcinoma) 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

  • Overlay histogram showing Jurkat (human T cell leukemia cell line from peripheral blood) 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.

  • All lanes : Anti-PARP1 antibody [E102] (ab32138) at 1/1000 dilution

    Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
    Lane 2 : Jurkat + Camptothecin cell lysate

    Predicted band size: 113 kDa
    Observed band size: 120,25 kDa
    why is the actual band size different from the predicted?

References

This product has been referenced in:
  • Zhang K  et al. PU.1/microRNA-142-3p targets ATG5/ATG16L1 to inactivate autophagy and sensitize hepatocellular carcinoma cells to sorafenib. Cell Death Dis 9:312 (2018). Read more (PubMed: 29472524) »
  • Palazzo L  et al. Serine is the major residue for ADP-ribosylation upon DNA damage. Elife 7:N/A (2018). Read more (PubMed: 29480802) »
See all 33 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (THP1 Cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin gel, semi dry transfer)
Loading amount
50 µg
Specification
THP1 Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jul 27 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Kidney)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin)
Loading amount
56 µg
Specification
Kidney
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 May 16 2013

Question
Answer

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.

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

Dr. Kirk Mcmanus

Verified customer

Submitted 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)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
No
Specification
Intestine
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 16 2012

Answer

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.

Read More

Answer

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 cleavage recognition sequences. Their experiments show MMP-2 degradation products of PARP corresponding to those theoretically predicted. Western blots for PARP after MMP-2 cleavage reveal loss of PARP as well as appearance of a <48 kDa product. These products appear in a concentration-dependent manner along with a corresponding loss of the 116 kDa PARP full length protein band. FASEB Journal. 2004;18:690-692.) Matrix metalloproteinase-2 (MMP-2) is present in the nucleus of cardiac myocytes and is capable of cleaving poly (ADP-ribose) polymerase (PARP) in vitro. Jennifer Kwan et al. At website address: http://www.fasebj.org/cgi/content/full/18/6/690?ck=nck I hope this helps to explain your results. I can suggest doing a further literature to obtain more information on this. If you are still concerned about the results you have obtained, please do not hesitate to complete the following questionnaire. This will provide me with your protocol information so I can investigate this case further. https://www.abcam.com/index.html?section=western&pageconfig=technical&mode=questionaire If you have any further questions, please do not hesitate to contact us again. Good luck with your experiments.

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