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BD Horizon™ BV421 Mouse Anti-Cleaved PARP (Asp 214)

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BD Horizon™ BV421 Mouse Anti-Cleaved PARP (Asp 214)

品牌： BD Pharmingen

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参数

反应种属：

Human (QC Testing), Mouse (Tested in Development)

来源宿主：

Mouse IgG1, κ

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产品介绍

产品信息

耦联标记

BV421

抗原名称

Cleaved PARP

宿主

Mouse IgG1, κ

免疫原

Human cleaved PARP

简单描述

PARP (Poly [ADP-Ribose] Polymerase) is a 113-kDa nuclear chromatin-associated enzyme that catalyzes the transfer of ADP-ribose units from NAD + to a variety of nuclear proteins including topoisomerases, histones, and PARP itself. The catalytic activity of PARP is increased in cells following DNA damage, and PARP is thought to play an important role in mediating the normal cellular response to DNA damage. Additionally, PARP is a target of the caspase protease activity associated with apoptosis. The PARP protein consists of an N-terminal DNA-binding domain (DBD) and a C-terminal catalytic domain separated by a central automodification domain. During apoptosis, Caspase-3 cleaves PARP at a recognition site (Asp Glu Val Asp Gly) in the DBD to form 24- and 89-kDa fragments. This process separates the DBD (which is mostly in the 24-kDa fragment) from the catalytic domain (in the 89-kDa fragment) of the enzyme, resulting in the loss of normal PARP function. It has been proposed that inactivation of PARP directs DNA-damaged cells to undergo apoptosis rather than necrotic degradation, and the presence of the 89-kDa PARP cleavage fraction is considered to be a marker of apoptosis. A peptide corresponding to the N-terminus of the cleavage site (Asp 214) of human PARP was used as the immunogen. The F21-852 monoclonal antibody reacts only with the 89-kDa fragment of human PARP-1 that is downstream of the Caspase-3 cleavage site (Asp214) and contains the automodification and catalytic domains. It does not react with intact human PARP-1. Cross-reactivity with other members of the PARP superfamily is unknown. Recognition of cleaved PARP in mouse cells has been demonstrated, and it may also cross-react with a number of other species due to the conserved nature of the molecule.

商品描述

F21-852 PARP (Poly [ADP-Ribose] Polymerase) is a 113-kDa nuclear chromatin-associated enzyme that catalyzes the transfer of ADP-ribose units from NAD + to a variety of nuclear proteins including topoisomerases, histones, and PARP itself. The catalytic activity of PARP is increased in cells following DNA damage, and PARP is thought to play an important role in mediating the normal cellular response to DNA damage. Additionally, PARP is a target of the caspase protease activity associated with apoptosis. The PARP protein consists of an N-terminal DNA-binding domain (DBD) and a C-terminal catalytic domain separated by a central automodification domain. During apoptosis, Caspase-3 cleaves PARP at a recognition site (Asp Glu Val Asp Gly) in the DBD to form 24- and 89-kDa fragments. This process separates the DBD (which is mostly in the 24-kDa fragment) from the catalytic domain (in the 89-kDa fragment) of the enzyme, resulting in the loss of normal PARP function. It has been proposed that inactivation of PARP directs DNA-damaged cells to undergo apoptosis rather than necrotic degradation, and the presence of the 89-kDa PARP cleavage fraction is considered to be a marker of apoptosis. A peptide corresponding to the N-terminus of the cleavage site (Asp 214) of human PARP was used as the immunogen. The F21-852 monoclonal antibody reacts only with the 89-kDa fragment of human PARP-1 that is downstream of the Caspase-3 cleavage site (Asp214) and contains the automodification and catalytic domains. It does not react with intact human PARP-1. Cross-reactivity with other members of the PARP superfamily is unknown. Recognition of cleaved PARP in mouse cells has been demonstrated, and it may also cross-react with a number of other species due to the conserved nature of the molecule.

同种型

Mouse IgG1, κ

克隆号

克隆 F21-852 (RUO)

产品详情

BV421

The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

格式:BV421

激发源:Violet 405 nm

最多激发源:407 nm

最多发射源:423 nm

应用

实验应用

Intracellular staining (flow cytometry) (Routinely Tested), Immunocytochemistry, Immunofluorescence (Tested During Development)

推荐用量

5 µl

反应种属

Human (QC Testing), Mouse (Tested in Development)

目标/特异性

Cleaved PARP (Asp214)

背景

别名

Asp214

翻译后修饰

Cleaved

制备和贮存

存储溶液

Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

保存方式

Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

文献

研发参考(10) 1. Amé J-C, Spenlehauer C, de Murcia G. The PARP superfamily. Bioessays. 2004; 26:882-893. (Biology). 2. Boulares AH, Yakovlev AG, Ivanova V, et al. Role of Poly(ADP-ribose) polymerase (PARP) cleavage in apoptosis. Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells. J Biol Chem. 1999; 274(33):22932-22940. (Biology). 3. Cherney BW, McBride OW, Chen D, et al. cDNA sequence, protein structure, and chromosomal location of the human gene for poly(ADP-ribose) polymerase. Proc Natl Acad Sci U S A. 1987; 84(23):8370-8374. (Biology). 4. D'Amours D, Desnoyers S, D'Silva I, Poirier GG. Poly(ADP-ribosyl)ation reactions in the regulation of nucelar functions. Biochem J. 1999; 342:249-268. (Biology). 5. Kaufmann SH, Desnoyers S, Ottaviano Y, Davidson NE, Poirier GG. Specific proteolytic cleavage of poly(ADP-ribose) polymerase: an early marker of chemotherapy-induced apoptosis. Cancer Res. 1993; 53(17):3976-3985. (Biology). 6. Lamarre D, Talbot B, Leduc Y, Muller S, Poirier G. Production and characterization of monoclonal antibodies specific for the functional domains of poly(ADP-ribose) polymerase. Biochem Cell Biol. 1986; 64(4):368-376. (Biology). 7. Lamarre D, Talbot B, de Murcia G, et al. Structural and functional analysis of poly(ADP ribose) polymerase: an immunological study. Biochim Biophys Acta. 1988; 950(2):147-160. (Biology). 8. Patel T, Gores GJ, Kaufmann SH. The role of proteases during apoptosis. FASEB J. 1996; 10(5):587-597. (Biology). 9. Soldani G, Scovassi AI. Poly(ADP-ribose) polymerase-1 cleavage during apoptosis: an update. Apoptosis. 2002; 7:321-328. (Biology). 10. Tewari M, Quan LT, O'Rourke K, et al. Yama/CPP32 beta, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. Cell. 1995; 81(5):801-809. (Biology).

数据库链接

Entrez-Gene ID

142, 11545

参考图片

Flow cytometric analysis of cleaved PARP (Asp 214) expression (left panel). Cells from the human Jurkat (ATCC TIB-152) cell line were cultured for 6 hours with (solid line histogram) or without (dashed line histogram) Camptothecin (Sigma-Aldrich Cat. No. C-9911; 12 μM final concentration). The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with BD Horizon™ BV421 Mouse Anti-Cleaved PARP (Asp 214) antibody (Cat. No. 564129). Histograms were derived from gated events with the forward and side light-scatter characteristics of intact Jurkat cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. Immunofluorescent analysis of cleaved PARP in apoptotic human cells (right panel). HeLa cells (ATCC, CCL-2) were treated with camptothecin (20 µM, 6 hours) to induce apoptosis. Cells were then fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050), and blocked with 5% goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in PBS. Cells were stained with BD Horizon™ BV421 Mouse Anti-Cleaved PARP (Asp 214) antibody (Cat. No. 564129, pseudo-colored red) and Alexa Fluor® 488 Mouse Anti-GM130 antibody (Cat. No. 560257, pseudo-colored green). DRAQ5 was used as a nuclear counterstain (Cat. No. 564902/564903, pseudo-colored blue). Images were captured on a standard epifluorescence microscope. Original magnification, 20x.

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货号：

564129

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询价

100Tst

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