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:10/11/2017)... ... October 11, 2017 , ... ... granted orphan drug designation to SBT-100, its novel anti-STAT3 (Signal Transducer and Activator ... osteosarcoma. SBT-100 is able to cross the cell membrane and bind intracellular STAT3 ...
(Date:10/10/2017)... ... October 10, 2017 , ... ... (ADC) therapeutics, today confirmed licensing rights that give it exclusive global access ... developed in collaboration with Children’s Hospital Los Angeles (CHLA). Additionally, an ...
(Date:10/10/2017)... ... , ... Dr. Bob Harman, founder and CEO of VetStem Biopharma, Inc. ... The event entitled “Stem Cells and Their Regenerative Powers,” was held on August ... MPVM was joined by two human doctors: Peter B. Hanson, M.D., Chief of Orthopedic ...
(Date:10/10/2017)... Calif. , Oct. 10, 2017 SomaGenics ... from the NIH to develop RealSeq®-SC (Single Cell), expected ... for profiling small RNAs (including microRNAs) from single cells ... Program highlights the need to accelerate development of approaches ... "New techniques for measuring levels ...
Breaking Biology Technology:
(Date:4/11/2017)... 11, 2017 Crossmatch®, a globally-recognized leader ... today announced that it has been awarded a ... Activity (IARPA) to develop next-generation Presentation Attack Detection ... "Innovation has been a driving force within Crossmatch ... allow us to innovate and develop new technologies ...
(Date:4/5/2017)... , April 5, 2017  The Allen Institute for ... Cell Explorer: a one-of-a-kind portal and dynamic digital window ... imaging data, the first application of deep learning to ... stem cell lines and a growing suite of powerful ... for these and future publicly available resources created and ...
(Date:3/30/2017)... , March 30, 2017  On April 6-7, 2017, ... the Genome hackathon at Microsoft,s headquarters in ... competition will focus on developing health and wellness apps ... Hack the Genome is the first hackathon ... The world,s largest companies in the genomics, tech and ...
Breaking Biology News(10 mins):