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Anti-PARP1 antibody [EPR18461] ab191217 is a rabbit monoclonal antibody that is used in PARP1 western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with PARP1 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR18461 is cited in over 120 publications

New 20 ul size available


Images

Western blot - Anti-PARP1 antibody [EPR18461] (AB191217), expandable thumbnail
  • Western blot - Anti-PARP1 antibody [EPR18461] (AB191217), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] (AB191217), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] (AB191217), expandable thumbnail
  • Western blot - Anti-PARP1 antibody [EPR18461] (AB191217), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IFIHC-P
Human
Tested
Tested
Tested
Mouse
Tested
Tested
Tested
Rat
Tested
Expected
Tested

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Rat

Dilution info

1/1000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Human

Dilution info

1/1000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species

Mouse

Dilution info

1/500

Notes

-

Species

Human

Dilution info

1/500

Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Rat

Dilution info

1/1000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Human

Dilution info

1/1000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

Function

Poly-ADP-ribosyltransferase that mediates poly-ADP-ribosylation of proteins and plays a key role in DNA repair (PubMed:17177976, PubMed:18055453, PubMed:18172500, PubMed:19344625, PubMed:19661379, PubMed:20388712, PubMed:21680843, PubMed:22582261, PubMed:23230272, PubMed:25043379, PubMed:26344098, PubMed:26626479, PubMed:26626480, PubMed:30104678, PubMed:31796734, PubMed:32028527, PubMed:32241924, PubMed:32358582, PubMed:33186521, PubMed:34465625, PubMed:34737271). Mediates glutamate, aspartate, serine, histidine or tyrosine ADP-ribosylation of proteins: the ADP-D-ribosyl group of NAD(+) is transferred to the acceptor carboxyl group of target residues and further ADP-ribosyl groups are transferred to the 2'-position of the terminal adenosine moiety, building up a polymer with an average chain length of 20-30 units (PubMed:19764761, PubMed:25043379, PubMed:28190768, PubMed:29954836, PubMed:35393539, PubMed:7852410, PubMed:9315851). Serine ADP-ribosylation of proteins constitutes the primary form of ADP-ribosylation of proteins in response to DNA damage (PubMed:33186521, PubMed:34874266). Specificity for the different amino acids is conferred by interacting factors, such as HPF1 and NMNAT1 (PubMed:28190768, PubMed:29954836, PubMed:32028527, PubMed:33186521, PubMed:33589610, PubMed:34625544, PubMed:34874266). Following interaction with HPF1, catalyzes serine ADP-ribosylation of target proteins; HPF1 confers serine specificity by completing the PARP1 active site (PubMed:28190768, PubMed:29954836, PubMed:32028527, PubMed:33186521, PubMed:33589610, PubMed:34625544, PubMed:34874266). Also catalyzes tyrosine ADP-ribosylation of target proteins following interaction with HPF1 (PubMed:29954836, PubMed:30257210). Following interaction with NMNAT1, catalyzes glutamate and aspartate ADP-ribosylation of target proteins; NMNAT1 confers glutamate and aspartate specificity (By similarity). PARP1 initiates the repair of DNA breaks: recognizes and binds DNA breaks within chromatin and recruits HPF1, licensing serine ADP-ribosylation of target proteins, such as histones (H2BS6ADPr and H3S10ADPr), thereby promoting decompaction of chromatin and the recruitment of repair factors leading to the reparation of DNA strand breaks (PubMed:17177976, PubMed:18172500, PubMed:19344625, PubMed:19661379, PubMed:23230272, PubMed:27067600, PubMed:34465625, PubMed:34874266). HPF1 initiates serine ADP-ribosylation but restricts the polymerase activity of PARP1 in order to limit the length of poly-ADP-ribose chains (PubMed:33683197, PubMed:34732825, PubMed:34795260). In addition to base excision repair (BER) pathway, also involved in double-strand breaks (DSBs) repair: together with TIMELESS, accumulates at DNA damage sites and promotes homologous recombination repair by mediating poly-ADP-ribosylation (PubMed:26344098, PubMed:30356214). Mediates the poly-ADP-ribosylation of a number of proteins, including itself, APLF, CHFR, RPA1 and NFAT5 (PubMed:17396150, PubMed:19764761, PubMed:24906880, PubMed:34049076). In addition to proteins, also able to ADP-ribosylate DNA: catalyzes ADP-ribosylation of DNA strand break termini containing terminal phosphates and a 2'-OH group in single- and double-stranded DNA, respectively (PubMed:27471034). Required for PARP9 and DTX3L recruitment to DNA damage sites (PubMed:23230272). PARP1-dependent PARP9-DTX3L-mediated ubiquitination promotes the rapid and specific recruitment of 53BP1/TP53BP1, UIMC1/RAP80, and BRCA1 to DNA damage sites (PubMed:23230272). PARP1-mediated DNA repair in neurons plays a role in sleep: senses DNA damage in neurons and promotes sleep, facilitating efficient DNA repair (By similarity). In addition to DNA repair, also involved in other processes, such as transcription regulation, programmed cell death, membrane repair, adipogenesis and innate immunity (PubMed:15607977, PubMed:17177976, PubMed:19344625, PubMed:27256882, PubMed:32315358, PubMed:32844745, PubMed:35124853, PubMed:35393539, PubMed:35460603). Acts as a repressor of transcription: binds to nucleosomes and modulates chromatin structure in a manner similar to histone H1, thereby altering RNA polymerase II (PubMed:15607977, PubMed:22464733). Acts both as a positive and negative regulator of transcription elongation, depending on the context (PubMed:27256882, PubMed:35393539). Acts as a positive regulator of transcription elongation by mediating poly-ADP-ribosylation of NELFE, preventing RNA-binding activity of NELFE and relieving transcription pausing (PubMed:27256882). Acts as a negative regulator of transcription elongation in response to DNA damage by catalyzing poly-ADP-ribosylation of CCNT1, disrupting the phase separation activity of CCNT1 and subsequent activation of CDK9 (PubMed:35393539). Involved in replication fork progression following interaction with CARM1: mediates poly-ADP-ribosylation at replication forks, slowing fork progression (PubMed:33412112). Poly-ADP-ribose chains generated by PARP1 also play a role in poly-ADP-ribose-dependent cell death, a process named parthanatos (By similarity). Also acts as a negative regulator of the cGAS-STING pathway (PubMed:32315358, PubMed:32844745, PubMed:35460603). Acts by mediating poly-ADP-ribosylation of CGAS: PARP1 translocates into the cytosol following phosphorylation by PRKDC and catalyzes poly-ADP-ribosylation and inactivation of CGAS (PubMed:35460603). Acts as a negative regulator of adipogenesis: catalyzes poly-ADP-ribosylation of histone H2B on 'Glu-35' (H2BE35ADPr) following interaction with NMNAT1, inhibiting phosphorylation of H2B at 'Ser-36' (H2BS36ph), thereby blocking expression of pro-adipogenetic genes (By similarity). Involved in the synthesis of ATP in the nucleus, together with NMNAT1, PARG and NUDT5 (PubMed:27257257). Nuclear ATP generation is required for extensive chromatin remodeling events that are energy-consuming (PubMed:27257257).Poly [ADP-ribose] polymerase 1, processed C-terminusPromotes AIFM1-mediated apoptosis (PubMed:33168626). This form, which translocates into the cytoplasm following cleavage by caspase-3 (CASP3) and caspase-7 (CASP7) in response to apoptosis, is auto-poly-ADP-ribosylated and serves as a poly-ADP-ribose carrier to induce AIFM1-mediated apoptosis (PubMed:33168626).Poly [ADP-ribose] polymerase 1, processed N-terminusThis cleavage form irreversibly binds to DNA breaks and interferes with DNA repair, promoting DNA damage-induced apoptosis.

Alternative names

ADPRT, PPOL, PARP1, PPOL, ADPRT, Poly [ADP-ribose] polymerase 1, PARP-1, ADP-ribosyltransferase diphtheria toxin-like 1, DNA ADP-ribosyltransferase PARP1, NAD(+) ADP-ribosyltransferase 1, Poly[ADP-ribose] synthase 1, Protein poly-ADP-ribosyltransferase PARP1, ARTD1, ADPRT 1

Recommended products

Anti-PARP1 antibody [EPR18461] ab191217 is a rabbit monoclonal antibody that is used in PARP1 western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with PARP1 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR18461 is cited in over 120 publications

New 20 ul size available

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR18461

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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 monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

PARP1 also known as poly(ADP-ribose) polymerase 1 is an enzyme that plays an important role in DNA repair processes. It detects DNA single-strand breaks and uses NAD+ as a substrate to add ADP-ribose polymers to itself and other proteins. This post-translational modification signals DNA repair machinery to the site of damage. PARP1 has a molecular weight of approximately 116 kDa. It is widely expressed in the nucleus of eukaryotic cells. PARP1 is often studied by western blotting techniques to analyze its expression and activation levels.

Biological function summary

Poly(ADP-ribose) polymerase 1 functions to maintain genomic stability by acting within the base excision repair complex. This complex is important for the detection and repair of DNA damage preventing the accumulation of mutations. By acting at sites of DNA stress PARP1 facilitates the binding of DNA repair proteins stabilizing the DNA structure during the repair process. This role is significant for cells that undergo frequent DNA replication or are exposed to high levels of genotoxic stress.

Pathways

The PARP1 protein is integral to the DNA damage response and repair pathway. It interacts with other proteins such as XRCC1 to coordinate repair activities at damaged DNA sites. Another important pathway involving PARP1 is the apoptosis pathway where excessive activation of PARP1 can lead to cell death due to depletion of cellular NAD+ and ATP. This indicates its dual role in both promoting cell survival through DNA repair and contributing to cell death when damage is irreparable.

Associated diseases and disorders

Poly(ADP-ribose) polymerase 1 is strongly linked to cancer and neurodegenerative diseases. Its activity is heightened in many cancer types where cancer cells exploit PARP1 for survival by repairing DNA damage that would otherwise be lethal. Inhibitors of PARP1 are being developed as cancer therapies to target these survival mechanisms. Moreover overactivation of PARP1 in neurodegenerative disorders like Alzheimer's disease can lead to excessive energy consumption promoting neuronal cell damage. In these contexts PARP1 connects with proteins like BRCA1 in cancer or AIF in neurodegeneration illustrating its role in disease mechanisms.

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12 product images

  • Western blot - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Western blot - Anti-PARP1 antibody [EPR18461] (ab191217)

    The lysates were prepared in 1%SDS Hot lysis method.

    Blocking/diluting buffer & concentration: 5% NFDM/TBST

    Observed MW: 112 kDa

    All lanes: Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution

    Lane 1: Rat brain lysates prepared in RIPA lysis method at 15 µg

    Lane 2: Rat brain lysates prepared in 1%SDS Hot lysis method at 15 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

    Predicted band size: 113 kDa

  • Western blot - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Western blot - Anti-PARP1 antibody [EPR18461] (ab191217)

    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 - ab191217 observed at 125 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    ab191217 was shown to specifically react with PARP1 when PARP1 knockout samples were used. Wild-type and PARP1 knockout samples were subjected to SDS-PAGE. ab191217 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-PARP1 antibody [EPR18461] (ab191217)

    Predicted band size: 113 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] (ab191217)

    Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling PARP1 with ab191217 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

    Nucleus staining on epithelial cells and stromal cells of Human testis is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] (ab191217)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PARP1 with ab191217 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HeLa cell line. The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:-
    -ve control 1: ab191217 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

  • Western blot - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Western blot - Anti-PARP1 antibody [EPR18461] (ab191217)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID: 1536009).

    The lysates were prepared in 1%SDS Hot lysis method

    All lanes: Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/10000 dilution

    Lane 1: Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 10 µg

    Lane 2: HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 1uM staurosporine for 4 hours whole cell lysates at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/50000 dilution

    Predicted band size: 113 kDa

    Observed band size: 113 kDa, 89 kDa

    Exposure time: 5s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] (ab191217)

    Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling PARP1 with ab191217 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

    Nucleus staining on epithelial cells and stromal cells of rat testis is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] (ab191217)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cells) cells labeling PARP1 with ab191217 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (red).

    The negative controls are as follows:-
    -ve control 1: ab191217 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

  • Western blot - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Western blot - Anti-PARP1 antibody [EPR18461] (ab191217)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID: 1536009).

    The lysates were prepared in 1%SDS Hot lysis method

    All lanes: Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/10000 dilution

    Lane 1: Untreated NIH/3T3 (Mouse embryonic fibroblast cells) whole cell lysates at 10 µg

    Lane 2: NIH/3T3 (Mouse embryonic fibroblast cells) treated with 1uM staurosporine for 4 hours whole cell lysates at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Predicted band size: 113 kDa

    Observed band size: 113 kDa, 55 kDa, 89 kDa

    Exposure time: 3min

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] (ab191217)

    Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling PARP1 with ab191217 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

    Nucleus staining on epithelial cells and stromal cells of mouse testis is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Western blot - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Western blot - Anti-PARP1 antibody [EPR18461] (ab191217)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The lysates were prepared in 1%SDS Hot lysis method

    All lanes: Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution

    Lane 1: Human fetal heart lysate at 10 µg

    Lane 2: Human fetal kidney lysate at 10 µg

    Secondary

    All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

    Predicted band size: 113 kDa

    Observed band size: 113 kDa

    Exposure time: 15s

  • Western blot - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Western blot - Anti-PARP1 antibody [EPR18461] (ab191217)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The lysates were prepared in 1%SDS Hot lysis method

    All lanes: Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution

    All lanes: Mouse heart lysate at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/50000 dilution

    Predicted band size: 113 kDa

    Observed band size: 113 kDa

    Exposure time: 1min

  • Western blot - Anti-PARP1 antibody [EPR18461] (ab191217), expandable thumbnail

    Western blot - Anti-PARP1 antibody [EPR18461] (ab191217)

    Western blot: Anti-PARP1 antibody [EPR18461] (ab191217) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab191217 was shown to bind specifically to PARP1. A band was observed at 125 kDa in wild-type A549 cell lysates with no signal observed at this size in PARP1 knockout cell line Human PARP1 knockout A549 cell line ab276094 (knockout cell lysate Abcam Pools). To generate this image, wild-type and PARP1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution

    Lane 1: Wild-type A549 control staurosporine (0 uM, 72 h) cell lysate at 20 µg

    Lane 2: Wild-type A549 treated staurosporine (3 uM, 24 h) cell lysate at 20 µg

    Lane 3: PARP1 knockout A549 control staurosporine (0 uM, 72 h) cell lysate at 20 µg

    Lane 4: PARP1 knockout A549 treated staurosporine (3 uM, 24 h) cell lysate at 20 µg

    Lane 5: Empty cell lysate at 20 µg

    Lane 6: HAP1 Treated Staurosporine (2uM, 4h) cell lysate at 20 µg

    Lane 7: HAP1 Vehicle Control Staurosporine (0uM, 4h) cell lysate at 10 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

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