Cleaved PARP Recombinant Rabbit Monoclonal Antibody [SU0314] - BSA and Azide free
Catalog# HA750136
Cleaved PARP Recombinant Rabbit Monoclonal Antibody [SU0314] - BSA and Azide free
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WB
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IF-Cell
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IP
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Human
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ET1608-10
含抗保成分
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unconjugated
Overview
Product Name
Cleaved PARP Recombinant Rabbit Monoclonal Antibody [SU0314] - BSA and Azide free
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
Product Features
Form
Liquid
Concentration
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000
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IF-Cell
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1:100-1:250
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IP
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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
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
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of Cleaved PARP on different lysates with Rabbit anti-Cleaved PARP antibody (HA750136) 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 (HA750136) 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 (HA750136) 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 (HA750136) 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 (HA750136) 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 HA750136 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750136 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: HA750136 IP in Daudi cell lysate
Lane 3: Rabbit IgG instead of HA750136 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"
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