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Index / Biovis / Caspase_2 Substrate VDVAD_pNA200 assays /Product Detail : 1072-200 Caspase_2 Substrate VDVAD_pNA200 assays
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#1072-200 Caspase_2 Substrate VDVAD_pNA200 assays

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  Price : 349   EUR
397   USD
271   GBP
1469   Zloty
46806   JPY
2697   NOK
2890   SEK
395   CHF

Product name : Caspase_2 Substrate VDVAD_pNA200 assays

Catalog number : 1072-200

Quantity: 200 assays

Availability: Yes

Supplier name : Biovis

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More Details about

STORAGE: Store at –20°C, protected from light. Stable for 6 months
MOL. WEIGHT: 679.0
SEQUENCE: Ac-Val-Asp-Val-Ala-Asp-pNA
PURITY: >98% by HPLC analysis


DESCRIPTION:
Ready-to-use colorimetric substrate for caspase-2/Ich-1 and related caspases that recognize the amino acid sequence VDVAD. Caspase-2 and related caspase activity can be quantified by detection of free pNA after cleavage from the peptide substrate VDVAD-pNA at OD405 nm using a spectrophotometer or plate reader. The ready-to-use caspase substrate provides an economic alternative for researchers who perform large amount of caspase assays.


ASSAY PROCEDURE:
1. Induce apoptosis in cells by desired method. Concurrently incubate a control culture without induction.
2. Count cells and pellet 1-5 x 106 cells.
3. Resuspend cells in 50 μl of chilled Cell Lysis Buffer (Cat.# 1067-100) and incubate cells on ice for 10 minutes.
4. Centrifuge for 1 min in a microcentrifuge (10,000 x g).
5. Transfer supernatant (cytosolic extract) to a fresh tube and put on ice.
6. Assay protein concentration.
7. Dilute 50-200 μg protein to 50 μl Cell Lysis Buffer for each assay.
8. Add 50 μl of 2X Reaction Buffer (Cat.# 1068-20, -80) containing 10 mM DTT (Cat.# 1201-1) to each sample.
9. Add 5 μl of the 4 mM of VDVAD-pNA (200 M final conc.) and incubate at 37°C for 1-2 hour.
9. Read samples at 400- or 405-nm in a microtiter plate reader, or spectrophotometer using a 100-μl micro quartz cuvet (Sigma), or dilute sample to 1 ml with Dilution Buffer (Cat.#1066-100, -500) and using regular cuvet (note: Dilution of the samples proportionally decreases the reading).
You may also perform the entire assay directly in a 96-well plate. Fold-increase in caspase activity can be determined by comparing these results with the level of the uninduced control.
Note: Background reading from cell lysates and buffers should be subtracted from the readings of both induced and the uninduced samples before calculating fold increase in caspase activity.

 

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Pathways :
WP1077: cytochrome P450
WP1194: cytochrome P450
WP1219: cytochrome P450
WP1274: cytochrome P450
WP1390: cytochrome P450
WP958: cytochrome P450

Related Genes :
[ppnP C7431_10447] Pyrimidine/purine nucleoside phosphorylase (EC 2.4.2.1) (EC 2.4.2.2) (Adenosine phosphorylase) (Cytidine phosphorylase) (Guanosine phosphorylase) (EC 2.4.2.15) (Inosine phosphorylase) (Thymidine phosphorylase) (EC 2.4.2.4) (Uridine phosphorylase) (EC 2.4.2.3) (Xanthosine phosphorylase)
[nnrE nnrD C7431_10346] Multifunctional fusion protein [Includes: ADP-dependent (S)-NAD(P)H-hydrate dehydratase (EC 4.2.1.136) (ADP-dependent NAD(P)HX dehydratase); NAD(P)H-hydrate epimerase (EC 5.1.99.6) (NAD(P)HX epimerase)]
[cysG C7431_10838] Siroheme synthase [Includes: Precorrin-2 dehydrogenase (EC 1.3.1.76); Sirohydrochlorin ferrochelatase (EC 4.99.1.4); Uroporphyrinogen-III C-methyltransferase (Urogen III methylase) (EC 2.1.1.107) (SUMT) (Uroporphyrinogen III methylase) (UROM)]
[cysG C7431_103526] Siroheme synthase [Includes: Precorrin-2 dehydrogenase (EC 1.3.1.76); Sirohydrochlorin ferrochelatase (EC 4.99.1.4); Uroporphyrinogen-III C-methyltransferase (Urogen III methylase) (EC 2.1.1.107) (SUMT) (Uroporphyrinogen III methylase) (UROM)]
[glmU C7431_109182] Bifunctional protein GlmU [Includes: Glucosamine-1-phosphate N-acetyltransferase (EC 2.3.1.157); UDP-N-acetylglucosamine pyrophosphorylase (EC 2.7.7.23) (N-acetylglucosamine-1-phosphate uridyltransferase)]
[murE C7431_10215] UDP-N-acetylmuramoyl-L-alanyl-D-glutamate--2,6-diaminopimelate ligase (EC 6.3.2.13) (Meso-A2pm-adding enzyme) (Meso-diaminopimelate-adding enzyme) (UDP-MurNAc-L-Ala-D-Glu:meso-diaminopimelate ligase) (UDP-MurNAc-tripeptide synthetase) (UDP-N-acetylmuramyl-tripeptide synthetase)
[arnA C7431_10733] Bifunctional polymyxin resistance protein ArnA [Includes: UDP-4-amino-4-deoxy-L-arabinose formyltransferase (EC 2.1.2.13) (UDP-L-Ara4N formyltransferase) (ArnAFT); UDP-glucuronic acid oxidase, UDP-4-keto-hexauronic acid decarboxylating (EC 1.1.1.305) (UDP-glucuronic acid dehydrogenase) (ArnADH) (UDP-GlcUA decarboxylase)]
[rlmN C7431_106167] Dual-specificity RNA methyltransferase RlmN (EC 2.1.1.192) (23S rRNA (adenine(2503)-C(2))-methyltransferase) (23S rRNA m2A2503 methyltransferase) (Ribosomal RNA large subunit methyltransferase N) (tRNA (adenine(37)-C(2))-methyltransferase) (tRNA m2A37 methyltransferase)
[hmp C7431_106150] Flavohemoprotein (Flavohemoglobin) (Hemoglobin-like protein) (Nitric oxide dioxygenase) (NO oxygenase) (NOD) (EC 1.14.12.17)
[rbsK C7431_11532] Ribokinase (RK) (EC 2.7.1.15)
[thiI C7431_10478] tRNA sulfurtransferase (EC 2.8.1.4) (Sulfur carrier protein ThiS sulfurtransferase) (Thiamine biosynthesis protein ThiI) (tRNA 4-thiouridine synthase)
[prs C7431_101271] Ribose-phosphate pyrophosphokinase (RPPK) (EC 2.7.6.1) (5-phospho-D-ribosyl alpha-1-diphosphate) (Phosphoribosyl diphosphate synthase) (Phosphoribosyl pyrophosphate synthase) (P-Rib-PP synthase) (PRPP synthase) (PRPPase)
[asd C7431_10880] Aspartate-semialdehyde dehydrogenase (ASA dehydrogenase) (ASADH) (EC 1.2.1.11) (Aspartate-beta-semialdehyde dehydrogenase)
[pyrG C7431_103508] CTP synthase (EC 6.3.4.2) (Cytidine 5'-triphosphate synthase) (Cytidine triphosphate synthetase) (CTP synthetase) (CTPS) (UTP--ammonia ligase)
[eno C7431_103509] Enolase (EC 4.2.1.11) (2-phospho-D-glycerate hydro-lyase) (2-phosphoglycerate dehydratase)
[garL C7431_103501] 5-keto-4-deoxy-D-glucarate aldolase (KDGluc aldolase) (KDGlucA) (EC 4.1.2.20) (2-dehydro-3-deoxy-D-glucarate aldolase) (2-keto-3-deoxy-D-glucarate aldolase) (5-dehydro-4-deoxy-D-glucarate aldolase) (Alpha-keto-beta-deoxy-D-glucarate aldolase)
[rbsK C7431_106237] Ribokinase (RK) (EC 2.7.1.15)
[dapB C7431_10259] 4-hydroxy-tetrahydrodipicolinate reductase (HTPA reductase) (EC 1.17.1.8)
[nagZ C7431_10175] Beta-hexosaminidase (EC 3.2.1.52) (Beta-N-acetylhexosaminidase) (N-acetyl-beta-glucosaminidase)
[serS C7431_104455] Serine--tRNA ligase (EC 6.1.1.11) (Seryl-tRNA synthetase) (SerRS) (Seryl-tRNA(Ser/Sec) synthetase)
[rbsK C7431_102102] Ribokinase (RK) (EC 2.7.1.15)
[argD dapC C7431_10829] Acetylornithine/succinyldiaminopimelate aminotransferase (ACOAT) (DapATase) (Succinyldiaminopimelate transferase) (EC 2.6.1.11) (EC 2.6.1.17)
[argD dapC C7431_101307] Acetylornithine/succinyldiaminopimelate aminotransferase (ACOAT) (DapATase) (Succinyldiaminopimelate transferase) (EC 2.6.1.11) (EC 2.6.1.17)
[C7431_103403] dITP/XTP pyrophosphatase (EC 3.6.1.66) (Non-canonical purine NTP pyrophosphatase) (Non-standard purine NTP pyrophosphatase) (Nucleoside-triphosphate diphosphatase) (Nucleoside-triphosphate pyrophosphatase) (NTPase)
[galK C7431_104311] Galactokinase (EC 2.7.1.6) (Galactose kinase)
[ackA C7431_105288] Acetate kinase (EC 2.7.2.1) (Acetokinase)
[hldE C7431_103233] Bifunctional protein HldE [Includes: D-beta-D-heptose 1-phosphate adenylyltransferase (EC 2.7.7.70) (D-glycero-beta-D-manno-heptose 1-phosphate adenylyltransferase); D-beta-D-heptose 7-phosphate kinase (EC 2.7.1.167) (D-beta-D-heptose 7-phosphotransferase) (D-glycero-beta-D-manno-heptose-7-phosphate kinase)]
[cca C7431_103229] Multifunctional CCA protein [Includes: CCA-adding enzyme (EC 2.7.7.72) (tRNA-NT) (tRNA nucleotidyltransferase) (tRNA adenylyl-/cytidylyl-transferase) (tRNA CCA-pyrophosphorylase) (CCA tRNA nucleotidyltransferase); 2'-nucleotidase (EC 3.1.3.-); Phosphatase; 2',3'-cyclic phosphodiesterase (EC 3.1.4.-)]
[ilvC C7431_108207] Ketol-acid reductoisomerase (NADP(+)) (KARI) (EC 1.1.1.86) (Acetohydroxy-acid isomeroreductase) (AHIR) (Alpha-keto-beta-hydroxylacyl reductoisomerase)
[pfkA C7431_11238] ATP-dependent 6-phosphofructokinase (ATP-PFK) (Phosphofructokinase) (EC 2.7.1.11) (Phosphohexokinase)

Bibliography :
[27019748] Caspase-2 resides in the mitochondria and mediates apoptosis directly from the mitochondrial compartment.
[23796689] A label-free LC/MS/MS-based enzymatic activity assay for the detection of genuine caspase inhibitors and SAR development.
[21903398] Exploiting differences in caspase-2 and -3 S₂ subsites for selectivity: structure-based design, solid-phase synthesis and in vitro activity of novel substrate-based caspase-2 inhibitors.
[21828056] Structural and enzymatic insights into caspase-2 protein substrate recognition and catalysis.
[14580374] IL-1 beta is a better inducer of apoptosis in human fetal membranes than IL-6.
[12460989] A novel Apaf-1-independent putative caspase-2 activation complex.
[12147221] Bcl-2 blocks apoptosis caused by pierisin-1, a guanine-specific ADP-ribosylating toxin from the cabbage butterfly.