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Knockout Tested Rabbit Recombinant Monoclonal PARP1 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB, Flow Cyt (Intra) and reacts with Human samples.

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Images

Western blot - Anti-PARP1 antibody [E102] - BSA and Azide free (AB221923), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [E102] - BSA and Azide free (AB221923), expandable thumbnail
  • Western blot - Anti-PARP1 antibody [E102] - BSA and Azide free (AB221923), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [E102] - BSA and Azide free (AB221923), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-PARP1 antibody [E102] - BSA and Azide free (AB221923), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PICC/IFWBFlow Cyt (Intra)IP
Human
Tested
Tested
Tested
Tested
Not recommended

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

Existing as a 113 kDa nuclear protein, PARP1 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)

Tested
Tested

Species

Human

Dilution info

-

Notes

ab199376- Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Not recommended
Not recommended

Species

Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

12 products for Alternative Product

2 products for Alternative Version

Target data

Function

Poly-ADP-ribosyltransferase that mediates poly-ADP-ribosylation of proteins and plays a key role in DNA repair (PubMed:17177976, PubMed:18172500, PubMed:19344625, PubMed:19661379, PubMed:23230272, PubMed:25043379, PubMed:33186521, PubMed:32028527, PubMed:26344098). Mediates glutamate, aspartate, serine 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:7852410, PubMed:9315851, PubMed:19764761, PubMed:25043379, PubMed:28190768, PubMed:29954836). Serine ADP-ribosylation of proteins constitutes the primary form of ADP-ribosylation of proteins in response to DNA damage (PubMed:33186521). Mainly mediates glutamate and aspartate ADP-ribosylation of target proteins in absence of HPF1 (PubMed:19764761, PubMed:25043379). Following interaction with HPF1, catalyzes serine ADP-ribosylation of target proteins; HPF1 conferring serine specificity by completing the PARP1 active site (PubMed:28190768, PubMed:29954836, PubMed:33186521, PubMed:32028527). Also catalyzes tyrosine ADP-ribosylation of target proteins following interaction with HPF1 (PubMed:30257210, PubMed:29954836). 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, 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). 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-ribosyl)ation of a number of proteins, including itself, APLF and CHFR (PubMed:17396150, PubMed:19764761). 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). Acts as a regulator of transcription: positively regulates the transcription of MTUS1 and negatively regulates the transcription of MTUS2/TIP150 (PubMed:19344625). Plays a role in the positive regulation of IFNG transcription in T-helper 1 cells as part of an IFNG promoter-binding complex with TXK and EEF1A1 (PubMed:17177976). 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).

Alternative names

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, PARP1, PPOL, ADPRT

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal PARP1 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB, Flow Cyt (Intra) and reacts with Human samples.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

Yes

Clone number

E102

Purification technique

Affinity purification Protein A

Specificity

This antibody recognises both pro-form and p25 cleaved form of PARP1.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab221923 is the carrier-free version of Anti-PARP1 antibody [E102] ab32138..

Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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.

Product promise

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7 product images

  • Western blot - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923), expandable thumbnail

    Western blot - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923)

    pro-form: 116kDa; p25 caspases cleaved form: 25kDa; proteolysis cleaved fragments: 58kDa and 42kDa

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

    Lane 1: Untreated Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate

    Lane 2: Jurkat (Human T cell leukemia T lymphocyte) treated with 1μM staurosporine for 4 hours whole cell lysate

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 113 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923)

    This data was developed using Anti-PARP1 antibody [E102] ab32138, the same antibody clone in a different buffer formulation.
    Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PARP1 with purified Anti-PARP1 antibody [E102] ab32138 at 1/100 dilution (1.0 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% TritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Western blot - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923), expandable thumbnail

    Western blot - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PARP1 antibody [E102] ab32138).

    Lanes 1- 2: Merged signal (red and green). Green - Anti-PARP1 antibody [E102] ab32138 observed at 113 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

    Anti-PARP1 antibody [E102] ab32138 was shown to react with PARP1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human PARP1 knockout HEK-293T cell line ab266598 (knockout cell lysate Human PARP1 knockout HEK-293T cell lysate ab257017). Wild-type HEK-293T and PARP1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Anti-PARP1 antibody [E102] ab32138 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

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

    Lane 1: Wild-type HEK-293T cell lysate at 20 µg

    Lane 2: PARP1 knockout HEK-293T cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 113 kDa

    Observed band size: 113 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923)

    This data was developed using Anti-PARP1 antibody [E102] ab32138, the same antibody clone in a different buffer formulation.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling PARP1 with purified Anti-PARP1 antibody [E102] ab32138 at 1/200 dilution (0.51 μg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Flow Cytometry (Intracellular) - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923)

    This data was developed using Anti-PARP1 antibody [E102] ab32138, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling PARP1 with purified Anti-PARP1 antibody [E102] ab32138 at 1/20 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Western blot - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923), expandable thumbnail

    Western blot - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923)

    Western blot: Anti-PARP1 antibody [E102] (Anti-PARP1 antibody [E102] ab32138) 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, Anti-PARP1 antibody [E102] ab32138 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 [E102] (Anti-PARP1 antibody [E102] ab32138) 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.

  • Flow Cytometry (Intracellular) - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-PARP1 antibody [E102] - BSA and Azide free (ab221923)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PARP1 antibody [E102] ab32138).
    Flow cytometry overlay histogram showing wild-type Hap1 (green line) and PARP1 knockout Hap1 stained with Anti-PARP1 antibody [E102] ab32138 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-PARP1 antibody [E102] ab32138) (1x 106 in 100μl at 0.04 μg/ml (1/55750)) for 30min at 22°C.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

    Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type Hap1 - black line, PARP1 knockout Hap1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

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For this product, it's our understanding that no specific protocols are required. You can:

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