PARP1 Recombinant Rabbit Monoclonal Antibody [SU03-68] - BSA and Azide free
Catalog# HA750154
PARP1 Recombinant Rabbit Monoclonal Antibody [SU03-68] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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FC
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Human
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Mouse
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Rat
概述
产品名称
PARP1 Recombinant Rabbit Monoclonal Antibody [SU03-68] - BSA and Azide free
抗体类型
Recombinant Rabbit monoclonal Antibody
免疫原
Synthetic peptide within N-terminal human PARP1.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IF-Tissue, IHC-P, FC
分子量
Predicted band size: 113 kDa
阳性对照
HeLa cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, C6 cell lysate, PC-12 cell lysate, HeLa whole cell lysate, HeLa treated with 1μM staurosporine for 3 hours whole cell lysate, HeLa, human breast cancer tissue, human colon tissue, mouse colon tissue, rat colon tissue.
偶联
unconjugated
克隆号
SU03-68
产品特性
形态
Liquid
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
PBS (pH7.4).
亚型
IgG
纯化方式
Protein A affinity purified.
应用稀释度
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WB
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1:2,000
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IF-Cell
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1:500
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IF-Tissue
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1:1,000
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IHC-P
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1:4,000
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FC
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1:1,000
靶点
功能
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. Cao C et al. The long intergenic noncoding RNA UFC1, a target of MicroRNA 34a, interacts with the mRNA stabilizing protein HuR to increase levels of -catenin in HCC cells. Gastroenterology 148:415-26.e18 (2015).
2. Gao S et al. Ischemia-reperfusion injury of the retina is linked to necroptosis via the ERK1/2-RIP3 pathway. Mol Vis 20:1374-87 (2014).
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, nucleolus, chromosome.
别名
ADP ribosyltransferase (NAD+
poly (ADP ribose) polymerase) 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
展开图片
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Western blot analysis of PARP1 on different lysates with Rabbit anti-PARP1 antibody (HA750154) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: Jurkat cell lysate (15 µg/Lane)
Lane 3: NIH/3T3 cell lysate (15 µg/Lane)
Lane 4: C2C12 cell lysate (15 µg/Lane)
Lane 5: C6 cell lysate (15 µg/Lane)
Lane 6: PC-12 cell lysate (15 µg/Lane)
Predicted band size: 113 kDa
Observed band size: 113 kDa
Exposure time: 6 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 (HA750154) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution were 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. -
☑ Cell treatment (CT)
Western blot analysis of PARP1 on different lysates with Rabbit anti-PARP1 antibody (HA750154) 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: 113 kDa
Observed band size: 113/28 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 (HA750154) 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)
All lanes: Western blot analysis of PARP with anti-PARP1 antibody[SU03-68] (HA750154) at 1:500 dilution.
Lane 1/2: Wild-type Hela whole cell lysate (10 µg).
Lane 3/4: PARP knockdown Hela whole cell lysate (10 µg).
HA750154 was shown to specifically react with PARP in wild-type Hela cells. Weakened bands were observed when PARP knockdown samples were tested. Wild-type and PARP knockdown 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 (HA750154, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling PARP1 with Rabbit anti-PARP1 antibody (HA750154) at 1/500 dilution.
Cells were fixed in 100% methanol for 5 minutes, blocked with 2% normal goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-PARP1 antibody (HA750154) at 1/500 dilution in 2% normal goat serum overnight at 4 ℃. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, green) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/800 dilution. -
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-PARP1 antibody (HA750154) at 1/4,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750154) at 1/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-PARP1 antibody (HA750154) at 1/4,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750154) at 1/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-PARP1 antibody (HA750154) at 1/4,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750154) at 1/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-PARP1 antibody (HA750154) at 1/4,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750154) at 1/4,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of HeLa cells labeling PARP1.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750154, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
Application: IF-Tissue
Species: Human
Site: breast cancer
Sample: Paraffin-embedded section
Antibody concentration: 1/1,000
请注意: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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