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使用敲除细胞株进行验证重组RabMAb

重组Anti-PARP1抗体[EPR18461] - BSA and Azide free (ab222234)

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Western blot - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
  • Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
  • Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
  • Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18461] to PARP1 - BSA and Azide free
  • Suitable for: IHC-P, ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 568 Alexa Fluor® 647 Alexa Fluor® 750 Unconjugated

概述

  • 产品名称

    Anti-PARP1抗体[EPR18461] - BSA and Azide free
    参阅全部 PARP1 一抗

  • 描述

    兔单克隆抗体[EPR18461] to PARP1 - BSA and Azide free

  • 宿主

    Rabbit

  • 经测试应用

    适用于: IHC-P, ICC/IF, WBmore details

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: HeLa and NIH/3T3 whole cell lysates; Human fetal heart and fetal kidney lysates; Mouse heart lysate; Rat brain and heart lysates. IHC-P: Human, mouse and rat testis tissues. ICC/IF: HeLa and NIH/3T3 cells.

  • 常规说明

    ab222234 is the carrier-free version of ab191217.

    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.

    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 production
    For more information see here.

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

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab222234于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 113, 89, 55 kDa (predicted molecular weight: 113 kDa).
说明
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 113, 89, 55 kDa (predicted molecular weight: 113 kDa).

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

    see all

  • 别名

    • 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
    • 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 1 antibody
    • Poly(ADP ribose) polymerase antibody
    • poly(ADP ribose) synthetase antibody
    • poly(ADP ribosyl)transferase antibody
    • Poly(ADP-ribosyl)transferase antibody
    • Poly[ADP ribose] synthetase 1 antibody
    • Poly[ADP-ribose] synthase 1 antibody
    • PPOL antibody
    • sPARP 1 antibody
    • sPARP1 antibody
    see all

图片

  • Western blot - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
    Western blot - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
    All lanes : 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

    Performed under reducing conditions.

    Predicted band size: 113 kDa
    Observed band size: 125 kDa why is the actual band size different from the predicted?



    Western blot: Anti-PARP1 antibody [EPR18461] (ab191217) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (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 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)

    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) (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) (ab97051) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191217).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)

    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) (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) (ab97051) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191217).

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

  • Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
    Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)

    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) (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 (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 (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191217).

  • Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
    Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)

    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) (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 (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (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) (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191217).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)

    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) (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) (ab97051) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191217).

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

  • Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)
    Anti-PARP1 antibody [EPR18461] - BSA and Azide free (ab222234)

文献 (0)

发表研究结果有使用 ab222234？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab222234 尚未被引用在任何文献中。

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Immunohistochemistry (Frozen sections) abreview for Anti-PARP1 antibody [EPR18461] - BSA and Azide free

 Inconclusive
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Renal Cortex)
Permeabilization
Yes - 0.3% Triton X-100
Specification
Renal Cortex
Blocking step
TSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: 20°C
Fixative
10 min Acetone (at room temperature) and 10 min 1.6% Paraformaldehyde (at room temperature) with a hydration step in PBS for 2 x 2 min in between
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Jan 30 2024

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