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Cleaved PARP Recombinant Rabbit Monoclonal Antibody [SU0314]

Usd: 385

Specification

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Catalog# ET1608-10

Cleaved PARP Recombinant Rabbit Monoclonal Antibody [SU0314]

Application

  • WB

  • IF-Cell

  • IP

Reactivity

  • Human

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Cleaved PARP Recombinant Rabbit Monoclonal Antibody [SU0314]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human PARP1 aa 200-249 / 1,014.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, IP

Molecular Weight

Predicted band size: 89 kDa

Positive Control

Jurkat cell lysate, A549 cell lysate, Hela, Daudi cell lysates.

Conjugation

unconjugated

Clone Number

SU0314

RRID

AB_3069785

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:100-1:250

  • IP

  • 1-2μg/sample

Target

Function

Poly (ADP-ribose) polymerase (PARP) is a family of proteins involved in a number of cellular processes such as DNA repair, genomic stability, and programmed cell death. The main role of PARP (found in the cell nucleus) is to detect and initiate an immediate cellular response to metabolic, chemical, or radiation-induced single-strand DNA breaks (SSB) by signaling the enzymatic machinery involved in the SSB repair. Once PARP detects a SSB, it binds to the DNA, undergoes a structural change, and begins the synthesis of a polymeric adenosine diphosphate ribose (poly (ADP-ribose) or PAR) chain, which acts as a signal for the other DNA-repairing enzymes. Target enzymes include DNA ligase III (LigIII), DNA polymerase beta (polβ), and scaffolding proteins such as X-ray cross-complementing gene 1 (XRCC1). After repairing, the PAR chains are degraded via Poly(ADP-ribose) glycohydrolase (PARG). PARP enzymes are essential in a number of cellular functions, including expression of inflammatory genes: PARP1 is required for the induction of ICAM-1 gene expression by cardiac myocytes and smooth muscle cells, in response to TNF.

Background References

1. Peh J et al. The Combination of Vemurafenib and Procaspase-3 Activation Is Synergistic in Mutant BRAF Melanomas. Mol Cancer Ther 15:1859-69 (2016).

2. Gao L et al. Glycyrrhizic acid alleviates bleomycin-induced pulmonary fibrosis in rats. Front Pharmacol 6:215 (2015).

Post-translational Modification

Phosphorylated by PRKDC and TXK.; Poly-ADP-ribosylated on glutamate and aspartate residues by autocatalysis. Poly-ADP-ribosylated by PARP2; poly-ADP-ribosylation mediates the recruitment of CHD1L to DNA damage sites. ADP-ribosylated on serine by autocatalysis; serine ADP-ribosylation takes place following interaction with HPF1.; S-nitrosylated, leading to inhibit transcription regulation activity.

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P09874 Human

Synonyms

ADP-ribosyltransferase diphtheria toxin-like 1 antibody

ADPRT 1 antibody

ADPRT antibody

ADPRT1 antibody

APOPAIN antibody

ARTD1 antibody

NAD(+) ADP-ribosyltransferase 1 antibody

PARP antibody

PARP-1 antibody

PARP1 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Cleaved PARP on different lysates with Rabbit anti-Cleaved PARP antibody (<a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa whole cell lysate<br />Lane 2: HeLa treated with 1μM staurosporine for 3 hours whole cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 89 kDa<br />Observed band size: 89 kDa<br /><br />Exposure time: 1 minute 9 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Cleaved PARP on different lysates with Rabbit anti-Cleaved PARP antibody (ET1608-10) at 1/2,000 dilution.

    Lane 1: HeLa whole cell lysate
    Lane 2: HeLa treated with 1μM staurosporine for 3 hours whole cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 89 kDa
    Observed band size: 89 kDa

    Exposure time: 1 minute 9 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-10) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Cleaved PARP on different lysates with Rabbit anti-Cleaved PARP antibody (<a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a>) at 1/500 dilution.<br /><br />Lane 1: Hela-si NT cell lysate (10 µg/Lane)<br />Lane 2: Hela-si Cleaved PARP cell lysate (10 µg/Lane)<br /><br />Predicted band size: 89 kDa<br />Observed band size: 89 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br /><a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a> was shown to specifically react with Cleaved PARP in Hela-si NT cells. No band was observed when Hela-si Cleaved PARP sample was tested. Hela-si NT and Hela-si Cleaved PARP samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a>, 1/500) and Loading control antibody (Rabbit anti-GAPDH, <a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Cleaved PARP on different lysates with Rabbit anti-Cleaved PARP antibody (ET1608-10) at 1/500 dilution.

    Lane 1: Hela-si NT cell lysate (10 µg/Lane)
    Lane 2: Hela-si Cleaved PARP cell lysate (10 µg/Lane)

    Predicted band size: 89 kDa
    Observed band size: 89 kDa

    Exposure time: 3 minutes;

    4-20% SDS-PAGE gel.

    ET1608-10 was shown to specifically react with Cleaved PARP in Hela-si NT cells. No band was observed when Hela-si Cleaved PARP sample was tested. Hela-si NT and Hela-si Cleaved PARP samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1608-10, 1/500) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of HeLa cells treated with 1μM staurosporine for 180 minutes labeling Cleaved PARP with Rabbit anti-Cleaved PARP antibody (<a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a>) at 1/250 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cleaved PARP antibody (<a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a>) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of HeLa cells treated with 1μM staurosporine for 180 minutes labeling Cleaved PARP with Rabbit anti-Cleaved PARP antibody (ET1608-10) at 1/250 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cleaved PARP antibody (ET1608-10) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Cleaved PARP was immunoprecipitated from 0.2 mg Daudi cell lysate with <a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a> at 1/500 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: Daudi cell lysate (input)<br />Lane 2: <a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a> IP in Daudi cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1608-10" style="font-weight: bold;text-decoration: underline;">ET1608-10</a> in Daudi cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 12 seconds; ECL: K1801

    Cleaved PARP was immunoprecipitated from 0.2 mg Daudi cell lysate with ET1608-10 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1608-10 at 1/500 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: Daudi cell lysate (input)
    Lane 2: ET1608-10 IP in Daudi cell lysate
    Lane 3: Rabbit IgG instead of ET1608-10 in Daudi cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 12 seconds; ECL: K1801

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

Citation

Products with the same target and pathway

metafield image

Cleaved PARP Recombinant Rabbit Monoclonal Antibody [SU0314] - BSA and Azide free

Application: WB,IF-Cell,IP

Reactivity: Human

Conjugate: unconjugated