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Index / Trevigen / Universal Chemiluminescent PARP Assay Kit w_ Histone Coated Strip wells / Product Detail : 4676-096-K Universal Chemiluminescent PARP Assay Kit w_ Histone Coated Strip wells
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Dec
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#4676-096-K Universal Chemiluminescent PARP Assay Kit w_ Histone Coated Strip wells

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  Price : 589   EUR
669   USD
457   GBP
2476   Zloty
78870   JPY
4545   NOK
4870   SEK
666   CHF

Product name : Universal Chemiluminescent PARP Assay Kit w_ Histone Coated Strip wells

Catalog number : 4676-096-K

Quantity: 96 Samples

Availability: Yes

Supplier name : Trevigen

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More Details about

I. Introduction
Poly ADP-ribosylation of nuclear proteins is a post-translational event that occurs in response to DNA damage. Poly (ADP-ribose) polymerase (PARP) catalyzes the NAD-dependent addition of poly (ADP-ribose) to itself and adjacent nuclear proteins such as histones. PARP contributes to the sequence of events that occurs during DNA base excision repair. Whereas PARP-mediated induction of necrosis can occur by extensive depletion of the intracellular NAD pool, the cleavage of PARP- promotes apoptosis by preventing DNA repair-induced survival and by blocking energy depletion-induced necrosis.Experimental models have shown that PARP inhibition prevents tissue damage in animal models of myocardial and neuronal ischemia, diabetes, septic shock, and vascular stroke. Moreover, PARP inhibition promotes chemosensitization and radiosensitization of tumors. Trevigen's HT Universal 96-well PARP Assay Kits measure the incorporation of biotinylated poly(ADP-ribose) onto histone proteins in a strip well format. This assay is ideal for the screening of PARP inhibitors and determining IC50 values.


Important features of the assay include:
1) chemiluminescent, non-radioactiveformat;
2) higher throughput 96 test size; and
3) sensitivity down to 0.01 Units of PARP per well. Histone-coated 96-well white strip wells (Cat# 4678-096-P) are available separately for your convenience.


II. Precautions and Limitations
1. For Research Use Only. Not for use in diagnostic procedures.
2. The physical, chemical, and toxicological properties of the chemicals and reagents contained in the HT Universal Chemiluminescent PARP Assay Kit may not yet have been fully investigated. Therefore, Trevigen recommends the use of gloves, lab coats, and eye protection while using any of these chemical reagents. Trevigen assumes no liability for damage resulting from handling or contact with these products. MSDS are available on request.

IV. Materials/Equipment Required But Not Supplied


Reagents:
1. Inhibitors
2. PBS (4870-500)
3. PBS + 0.1% Triton X-100
4. Distilled water

Disposables:
1. 1 - 200 μl and 100-1000 μl pipette tips


Equipment:
1. Micropipettes
2. Multichannel pipettor 10 - 200 μl
3. Wash bottle or microstrip wells washer (optional)
4. 96-well plate reader


V. Reagent Preparation
1. 10X Strep-Diluent
This solution is used as a diluent for the Strep-HRP. Dilute 1:10 with dH20 before use.
2. 20X PARP Buffer
Dilute the 20X PARP Buffer to 1X (1:20) with dH2O. The 1X PARP Buffer is used to rehydrate the histone coated wells, and to dilute the enzyme, PARP Cocktail, and the inhibitors to be tested.
3. 10X PARP Cocktail
Dilute the 10X PARP Cocktail as follows:
10X PARP Cocktail (Cat# 4671-096-03) 2.5 μl/well
10X Activated DNA (Cat# 4671-096-06) 2.5 μl/well
1X PARP Buffer 20 μl/well
4. PARP Enzyme
The kit contains 50 μl of PARP-HSA enzyme at a concentration described in the enclosed Product Data Sheet. The enzyme should be diluted appropriately with 1X PARP Buffer just before use. Note: Diluted enzyme should be used immediately and any remainder discarded.
5. PARP Inhibitors
The 3-aminobenzamide (3-AB) is provided at 200 mM in ethanol as a control inhibitor. 3-AB will inhibit the activity of PARP at a wide range of concentrations from 2 μM to 10 mM. Serially dilute the stock 3-AB or your PARP inhibitor(s) with 1X PARP Buffer and add to designated wells.
6. Strep-HRP
Just before use, dilute Strep-HRP (cat# 4800-30-06) 500-fold with 1X Strep-Diluent (cat# 4671-096-04). A total of 50 μl/well of diluted Strep-HRP is required in the assay.
7. PeroxyGlow™ A and B Chemiluminescent Substrates
Just before use mix equal volumes of PeroxyGlow™ A and B together. PeroxyGlow™ A and B are Horseradish Peroxidase (HRP) substrates generating light that can be quantified in a suitable chemiluminescent strip wells reader.

 

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