Navigation Links
A Further Step in Understanding Apoptosis,,, Direct Detection of PARP Cleavage

Detection of early apoptotic events has become quite relevant since many studies have demonstrated that abnormalities in cell death regulation cause a variety of diseases and pathological conditions accompanied either by cell accumulation (e.g., cancer) or by cell loss (e.g., Alzheimers dementia).

One of the key initiation elements of the apoptotic pathway is the activation of caspases (cysteinyl-aspartic acid proteases) followed by cleavage of the caspase substrates [1]. The 113-kDa poly(ADP-ribose) polymerase 1 (PARP-1), which is normally involved in DNA repair, DNA stability, and other cellular events, is cleaved by members of the caspase family during early apoptosis. The zincdependent eukaryotic DNA-binding protein PARP-1 specifically recognizes DNA strand breaks produced by various genotoxic agents. Detection of a 89-kDa or 24-kDa caspase cleavage fragment of PARP-1 was shown to be a hallmark of apoptosis [2]. In contrast to measuring active caspase 3, which is degraded during apoptosis via the ubiquitin/proteosome pathway, measuring PARP-1 cleavage might allow sustained signal detection even in late stages of apoptosis.

Product Description

With the anti-PARP Caspase Cleavage Product Antibody you will specifically detect only the small apoptotic PARP-1 cleavage fragment, but not the full-length protein, in samples of human, mouse, or rat origin. The fluorescein- conjugated monoclonal antibody (clone PAR-16) enables direct detection of this early apoptosis marker.

Furthermore it provides a tool for apoptotic-pathway analysis (in situ) at the stage of caspase activity. It can be easily employed in many different applications such as flow cytometry, immunocytochemistry, and immunohistochemistry (also in conjunction with an anti-fluorescein antibody). Particularly, in regard to tissue sections, the antibody is a valuable tool in acquiring additional information on the results achieved using the TUNEL method (even in dual-labeling applications).

Typical Experiment
Flow cytometric analysis of PARP cleavage and TUNEL reactivity

HeLa cells were induced to undergo apoptosis via stimulation with anti-CD95 (clone 2R2, 0.2 g/ml) and cycloheximide (10 g/ml). After 5 hours, the untreated cells (Figure 1, left panel) and CD95-stimulated cells (Figure 1, right panel) were stained for PARP cleavage using the anti-PARP Caspase Cleavage Product-Fluorescein antibody (1 g/ml), and analyzed for TUNEL staining using the In Situ Cell Death Detection Kit, TMR red.
Cells in the two right boxes of the dot-blot analysis display PARP-1 cleavage; the upper two boxes indicate TUNEL-positive cells (Figure 1, right panel). Note that PARP cleavage precedes DNA degradation and TUNEL labeling. The staining pattern of the apoptotic small PARP fragment in HeLa cells is also shown by immunofluorescence in Figure 2a. A peptide competition control experiment is included (Figure 2b).

To gain new insights into apoptotic pathways particularly in situ Roche Applied Science now offers the anti- PARP Caspase Cleavage Product-Fluorescein antibody. The antibody can serve as an early marker of apoptosis since it is directed against the caspase cleavage site of PARP-1 and specifically recognizes an epitope that is generated by caspases, but is not accessible in nonapoptotic cells.
This easy-to-use addition to the Roche caspase-signaling product line provides a number of benefits, including
  • Fast, simple detection in flow cytometry and fluorescence microscopy using a fluorescein-conjugate design
  • Sensitive technique for immunohistochemistry testing with tissue sections
  • Dual-labeling applications, such as combination with TUNEL
  • Apoptotic-pathway analysis (in situ) at the stage of caspase activation from early to later stages
  • Early apoptosis detection method in tissue sections complementing TUNEL data.


'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Counting Colonies or Plaques Directly from Images of Agar Plates
2. High-Efficiency Site-Directed Mutagenesis of Large Plasmid Vectors
3. Constructing Directional cDNA Libraries from Limited Amounts of RNA
4. Generate High-Quality, Directional Plasmid cDNA Libraries
5. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
6. Optimizing the Direct Amplification of Missing cDNA 5 Ends Using the Eppendorf Mastercycler gradient
7. TripleMaster PCR System: Direct Comparison with Competitors
8. DirectPrep 96 Miniprep System for cost-effective, high-throughput plasmid DNA purification
9. YOUR DATA: Direct Real-time RT-PCR on Sympathetic Neuron Lysates Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs
10. YOUR DATA: Direct Real-time RT-PCR on Sympathetic Neuron Lysates Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs
11. Determination of Polar Organophosphorus Pesticides in Aqueous Samples by Direct Injection Using HPLC/MS/MS
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/27/2016)... 2016  Sequenom, Inc. (NASDAQ: SQNM ), ... through the development of innovative products and services, announced ... United States denied its petition to review ... Sequenom,s U.S. Patent No. 6,258,540 (",540 Patent") are not ... the Supreme Court,s Mayo Collaborative Services v. Prometheus Laboratories ...
(Date:6/27/2016)... Raleigh, NC (PRWEB) , ... June 27, 2016 ... ... a mission to bring innovative medical technologies, services and solutions to the healthcare ... development and implementation of various distribution, manufacturing, sales and marketing strategies that are ...
(Date:6/27/2016)...   Ginkgo Bioworks , a leading organism ... today awarded as one of the World Economic ... most innovative companies. Ginkgo Bioworks is engineering biology ... world in the nutrition, health and consumer goods ... customers including Fortune 500 companies to design microbes ...
(Date:6/24/2016)... ... June 24, 2016 , ... While the majority of commercial spectrophotometers and ... and the 6000i models are higher end machines that use the more unconventional z-dimension ... light beam from the bottom of the cuvette holder. , FireflySci has developed ...
Breaking Biology Technology:
(Date:6/3/2016)... 3, 2016 Das ... Nepal hat ein 44 ... geprägter Kennzeichen, einschließlich Personalisierung, Registrierung und IT-Infrastruktur, ... Produktion und Implementierung von Identitätsmanagementlösungen. Zahlreiche renommierte ... Januar teilgenommen, aber Decatur wurde als konformste ...
(Date:5/24/2016)... Calif. , May 24, 2016 Ampronix facilitates superior patient care by ... LMD3251MT  3D medical LCD display is the latest premium product recently added to the ... ... ... Sony 3d Imaging- LCD Medical Display- Ampronix News ...
(Date:5/3/2016)... , May 3, 2016  Neurotechnology, a provider ... MegaMatcher Automated Biometric Identification System (ABIS) , ... multi-biometric projects. MegaMatcher ABIS can process multiple complex ... any combination of fingerprint, face or iris biometrics. ... SDK and MegaMatcher Accelerator , which ...
Breaking Biology News(10 mins):