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Index / Biovis / Caspase_5 Substrate WEHD_pNA200 assays /Product Detail : 1102-200 Caspase_5 Substrate WEHD_pNA200 assays
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#1102-200 Caspase_5 Substrate WEHD_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_5 Substrate WEHD_pNA200 assays

Catalog number : 1102-200

Quantity: 200 assays

Availability: Yes

Supplier name : Biovis

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More Details about

STORAGE: Store at –20°C, protected from light.
SHELF LIFE: 1 year under proper storage conditions
MOL. WEIGHT: 747.7
SEQUENCE: Ac-Trp-Glu-His-Asp-pNA
PURITY: >99% by HPLC analysis.


DESCRIPTION:
Ready-to-use colorimetric substrate for caspase-5 and related caspases that recognize the amino acid sequence WEHD. Caspase-5 and related caspase activity can be quantified by spectrophotometric detection of free pNA ( = 400 nm) after cleaved from the peptide substrate WEHD-pNA, using a spectrophotometer or multi-well plate reader. The ready-to-use caspase substrate provides an economic alternative for researchers who perform large volume 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 WEHD-pNA (200 μM final conc.) and incubate at 37°C for 1-2 hour.
10. 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 cuvette (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.


For Research Use Only! Not to be used in humans.

 

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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)]
[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)]
[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)]
[hmp C7431_106150] Flavohemoprotein (Flavohemoglobin) (Hemoglobin-like protein) (Nitric oxide dioxygenase) (NO oxygenase) (NOD) (EC 1.14.12.17)
[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)]
[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)
[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)
[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)
[rbsK C7431_11532] Ribokinase (RK) (EC 2.7.1.15)
[pyrG C7431_103508] CTP synthase (EC 6.3.4.2) (Cytidine 5'-triphosphate synthase) (Cytidine triphosphate synthetase) (CTP synthetase) (CTPS) (UTP--ammonia ligase)
[thiI C7431_10478] tRNA sulfurtransferase (EC 2.8.1.4) (Sulfur carrier protein ThiS sulfurtransferase) (Thiamine biosynthesis protein ThiI) (tRNA 4-thiouridine synthase)
[asd C7431_10880] Aspartate-semialdehyde dehydrogenase (ASA dehydrogenase) (ASADH) (EC 1.2.1.11) (Aspartate-beta-semialdehyde dehydrogenase)
[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)
[eno C7431_103509] Enolase (EC 4.2.1.11) (2-phospho-D-glycerate hydro-lyase) (2-phosphoglycerate dehydratase)
[serS C7431_104455] Serine--tRNA ligase (EC 6.1.1.11) (Seryl-tRNA synthetase) (SerRS) (Seryl-tRNA(Ser/Sec) synthetase)
[C7431_10469] Riboflavin biosynthesis protein RibD [Includes: Diaminohydroxyphosphoribosylaminopyrimidine deaminase (DRAP deaminase) (EC 3.5.4.26) (Riboflavin-specific deaminase); 5-amino-6-(5-phosphoribosylamino)uracil reductase (EC 1.1.1.193) (HTP reductase)]
[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)
[dusA C7431_102507] tRNA-dihydrouridine(20/20a) synthase (EC 1.3.1.-) (EC 1.3.1.91) (U20-specific dihydrouridine synthase) (U20-specific Dus) (tRNA-dihydrouridine synthase A)
[ilvC C7431_108207] Ketol-acid reductoisomerase (NADP(+)) (KARI) (EC 1.1.1.86) (Acetohydroxy-acid isomeroreductase) (AHIR) (Alpha-keto-beta-hydroxylacyl reductoisomerase)
[rbsK C7431_102102] Ribokinase (RK) (EC 2.7.1.15)
[nagZ C7431_10175] Beta-hexosaminidase (EC 3.2.1.52) (Beta-N-acetylhexosaminidase) (N-acetyl-beta-glucosaminidase)
[galK C7431_104311] Galactokinase (EC 2.7.1.6) (Galactose kinase)
[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)
[ackA C7431_105288] Acetate kinase (EC 2.7.2.1) (Acetokinase)
[pfkA C7431_11238] ATP-dependent 6-phosphofructokinase (ATP-PFK) (Phosphofructokinase) (EC 2.7.1.11) (Phosphohexokinase)

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