Cleaved PARP Recombinant Rabbit Monoclonal Antibody [SU0314]

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产品规格

50μl

100μl

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Catalog# ET1608-10

Cleaved PARP Recombinant Rabbit Monoclonal Antibody [SU0314]

Application

WB
IF-Cell
IP

Reactivity

Human

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

概述

产品名称

Cleaved PARP Recombinant Rabbit Monoclonal Antibody [SU0314]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

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

种属反应性

Human

验证应用

WB, IF-Cell, IP

分子量

Predicted band size: 89 kDa

阳性对照

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

偶联

unconjugated

克隆号

SU0314

RRID

AB_3069785

产品特性

形态

Liquid

存放说明

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.

存储缓冲液

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

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

WB
1:2,000

IF-Cell
1:100-1:250

IP
1-2μg/sample

发表文章中的应用

WB	查看 63 篇文献如下

发表文章中的种属

Human	查看 55 篇文献如下
Mouse	查看 13 篇文献如下
Rat	查看 2 篇文献如下
Monkey	查看 1 篇文献如下

靶点

功能

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.

背景文献

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).

翻译后修饰

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.

亚细胞定位

Nucleus.

UNIPROT

SwissProt: P09874 Human

别名

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

展开

图片

☑ 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.
☑ 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.
☑ 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 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