Chemiluminescent and chemifluorescent substrates produce a ‘more intense signal than colorimetric substrates, thus providing greater sensitivity. ELISA conditions must always be re-optimized ‘when switching toa diferent substrate. Greater dilations of @ conjugate is used witha chemiluminescent substrate than with a colorimetric substrate, Enzyme-abeled reagents are detected using chromogenic, chemiluminescent of cherflurescent substrates, hromagenie substrates are goneraly the least expensive, while Kminescent or ‘ton more sensive When perform ‘ng ELISAS a soluble substrates used and converted oa soluble snd product ELISAs are performed on polystyrene plates, andthe snzyme lavas are determined by monitoring signal development witha spectrophotometr [a luminometer for luminescence ora fluorometer for fluorescence) To order, call 0174-3728 o 15-56-0747, Outside the United Stats, contact your acl branch ofce or dnvibutrTle 6 Properties of Therme Scent ELISA Substrates horseradish peroxidase (HRP) alkaline p phatase (AP) and galactosidase (ab. Dittin ange ot 8 sebarte Protett Page Meserement Caer trom tg sick) Apri Sesiviy? Eye ‘SueSmnlSAfom CHC. MnmchneSCoR I~ ‘vpavel HP ‘Sipeina FSR Pio Ti) —«emchetnnecene OFpavel RP ‘amaBinSubiate SID «SSnmicamifcrscent 1° 1K= 120 (Segal HRP 2K 00K ‘manos ——=S~=*S*«USSSS CER chemminscent OK 12K Qiépgiet HRP k= 400K Petco Sp Ula oe ——_«Rem pga Yel T= 1K ‘awe ie Sar nrtop Be 2 aK 110 Perce Sap To39 TNS MOR —_«ONm copped Yel 11-1 Thawel HP ‘Sar nartop she erated Perce Sip Sow TS HOC ped Yolo T= 1K ‘pai 1h ‘Sar nertop she PAK 0 TW Sabet MO K«Stamtpped Yeo 00-1 Seve 1h ‘Sano nrtop Be ak ree Sap ABTS Tas «CMOS Geen 00-1 Cairne Tir ro GCOS een 00-1 Oairgnet 1 OPO Power «Aen Geen Ts Thawel RP ‘stro nnopevelbw-Orarse 2° HK 1900 OPO Ta a 1-1 Thaw oP {Soro nentop—elen-Orarge 24K 10K reap PNP ween —Yolow Tis Tora a Pisk FRPP Ke mane tse ‘ove co Esk PPP Oona ste ‘eae rs Esk RPP Power HOS inn Yolo ‘oes ro one ear Tae aa ‘Choosing a detection signal ype Coline Seas Chaiarascnt Sab Chonteineson Sabie Ts Medi erty ah sera ih sos Senerayes expense “Generally mre xsesve {ener expensive “Many subsets atte Fewscoeats sa SFanaibernes walle {Sh sgnlgenracn ‘Rape sant generon “Rast sil erereion Semele gly “Feeney mareaed Serereewace quetly 1 Spl rearrange onan way “lige Irearanglmhances card Fwaty “Lage 9 low-anéinerty “Foal cpp rnstoppe ards says) tppeoviopped angie assays] _* Nowe Deacon Faure ‘Soecwepoenr| ion, or to download product structions, isit wwwthermescientfc, mipierceHorseradish Peroxidase Substrates Horseradish peroxidase iso 4OkDa protein that catalyzes the ‘oxidation of substrates by hydrogen peroxide, esting in colored ‘or fluorescent product or the release of ight as a byproduct. HRP functions optimally ats near-neutal pH and can be nite by cyanides, suldes and azides, Antibody HRP conjugates are Superior to anabody-AP conjugates with respectto th specie actives ofboth the enzyme and antibody. In adeion, a high turnover rat, stably ow cost and wide aalabity of substrates make HRP the enqyme of ehoie or most applications When selecting a substrate for HRP-based ELISAs, chee ae a numberof posites, The question of sensitiv is oten the averting factor in making a selection. Howover, consideration Should also be given to whether te substrate contains harm Solvents what detection eouipmentis avaiable and how th willbe measured [kinetic o end-point In many ELISA applic tions, colorimetic substrates provide a suficiet love of sensitivity and dynamic range. This is evident with our cytokine ELISA k,n \wbich a TMB substrate combined wth streplaiin-HAP detection ‘system results in pg/mL sensitivity, Detection below this level requires a fluorescent or chemiluminescent signal. Characteristics of the HRP ELISA subsvates we ofa are summarized in Table 6 ay Chromogenic ELISA substrates result ina soluble, colored product, ‘These substales are used in mast FLISAs hecause detection af colored product ean be performed on a spectrophotomevie plate ir. TMB (3,35,5-tetramethylbenzidine) isthe most common ‘hromagenic substrate for HRP and is available in several formats ‘Thermo Scientific Pierce t=Step Ultra TMB (Product # 34028) yields ‘the greatest sonstivty amang the TMB substrates, fellawod by Pierce 1-Step Turbo TMB (Product # 24022) and Pierce T-Step Slow ‘TMB (Product #34024), The Pierce 1-Step Substrates are prefor mulated sono mixing or pre-fiterng is raqured. Although the sensitivity is lower, the Pierce 1-Stop Slaw TMB and Pierce T-Step ABTS (2,2-azinobis(S-ethybenzothiazoline-6-sulfonie acid]- iammonium salt are ideal for kinetic readings, OPD (o-phenyl ‘enediamine dihydrochloride, Product #24006) is another relatively sensitive HRP substrate that produces a yellow-orange color. ‘The greatest sensitivity in ELISA applications is obtained using ‘chamiluminascont or chomifiunrescont substrates These, substrates have been steadily qaining in popularity because oftheir sensitivity [ess than 1pq/ml} large linear range for detection and excellent antibody conservation. We offer the chemiluminescent Thormo Sciontfic SuporSignal ELISA Pico (Product #37070) and SuperSignal ELISA Femto (Product #27075) Substrates and the chemifluorescent Thermo Scientific QuantaBlu Substrate (Product #15169), sRate Baty. pvel oe we —o— ao When energy inthe form of lights released from a substance because of a chomical reaction, the process is called chomilum nescence, Lumina is one of the mast widely used chemilumines- ‘cent reagents and its oxidation by peroxide results in creation of an ‘excited state product called 3-aminophthalate, This product decays ‘to alower eneray state by releasing photons of ight (Figure 10} ~eag!.chak Figure 10, Lumiol is oxidized inthe presence of horseradish peroxidase rérogen peroxide to form an excited state product (-aminephthalats. “he Saminophthalate emits Ight ot 250m os deeays tothe ground state, To order, call 00174-5728 o 15-56-0747, Outside the United Stats, contact your acl branch ofce or dnb myChemiluminescent substrates have steadily gained in popularity ‘throughout the past decade because they offer several advantages var other detection mothods. Chemiluminascence has large linear rosponse for detection and quantitation aver a wide range of protein concentrations. Most importantly, chemiluminescence Yields the greatest sensitivity of any available detection method. 3 3 Chemiluminescent substrates differ from other substrates in that Thisis in contrast to substrates that produce a stable, colored product; these colors persist inthe well afer the enzyme-substrater has terminated tion ‘Awel-optimized procedure using the proper antibody dilutions will produce a stable output of ight, producing consistent and sensitve resuts. WRG This isthe single greatest cause of variability in chamiluminescont ELISAS. 908 of enhanc sentve ra Nonbacaeous oa at ate, te sha es eng iseeare Ress ae eased ig «ino HRP Substrates for ELISA Excellent sensitivity for use in luminometers, CChemiluminescent substrates are ideal for researchers looking for greater sensitivity in their ELISAs or any other solution-based assay. These ELISAS can take place in either a test tube or a microplate and are quantified by measuring relative light units {RLU) ina luminometer. Thermo Scientific SuperSignal ELISA Pico CChemiluminescent Substrate was developed for researchers who ‘need high sensitivity at an economical price. SuperSignal ELISA Fomto Maximum Sensitivity Chemiluminescent Substrate uses an ‘enhancer system that meets the needs of high-throughput screening and diagnostic. Both have their own unique features as listed in Table. Which substrate is ight Datacton subse Kina Ainaes Teme Seeatie —Weotewam lowed It Sipe SA tener Feet Maran Seo Subtle Stoatmince stably, ependng 0 HR cere Theme Seanle—plesyam lame ight Sips feneaton Eikapes neva abt Chanimiescent ua ‘SiberateSuperSignal ELISA Pico Chemiluminescent Substrate Get high sensitivity and a large dynamic range. ‘Thermo Scientific SuperSignal ELISA Pico Chemiluminescent Substrate is optimized fr luminometer-based assays to generate an intense signal. inate Analysis 20090 Bante RP 0 0 2 0 00 he ° e cy Tine nites —e— pease ee bans nin Working solitons of chemiluminescent subsvates were 38 aeeordng tothe manufacturers istuctons For SuperSignal ELISA Pico Cherluminescent Substrate and snatherlumnat-base system Brand A, WopL af each substrate working soluton was adaea tothe approrate plate wel For the doxetane-based sytem, wells were washed wth 1 Assay Buffer and TODpLef the dixetare working soliton was add tthe appropriate pate ‘Wo Alplates were nhated on a plate shaker arom tongeraure fT} 1 inate, Plates were then adeno plate uminrneter vith 2.2 secon read tine per wel. Several readings were taken aver a 3-ninute pig Highights: + Immediate generation of light intense immediately atroom temperature or at 37°C + High signalnoise ratio minimal background + Low picogram sensitivity detect proteins in your ELISAs down, tothe picagram levels + Room temperature storage ~@ consistent product with ambient shipping and no need to store at&°C + Bhour working solution stability - consistent performance af the ‘working solution over an &-hour period with only 10% decrease in activity at 24 hours + Flexible signal can be read in black ar white opaque plates + Emits fight at 425m als produced Aaleences Srdny A, a0 ial Chem a, coer, eke a 00), Genome or 156 Ordering Information Trio aniatEoarce sont. Sle oredr Ea To order, call 00174-5728 o 15-56-0747, Outside the United Stats, contact your acl branch ofce or dnb Ls |SuperSignal ELISA Femto Maximum Sensitivity Substrates Tho most powerful substrate for high-throughput screening/ diagnostic applications with high sensitviy and superior low-end linearity. Thermo Scientific SuperSignal ELISA Femte Maximum Senstvty Sub- strates formulated for superior protein detection and low-end linearity in ELISA applications, ‘SuperSigna ELISA Femto Substrate’ rapid signal generation has the benefit of decreasing the substrate incubation period tng detectable light within 1 minute, saving upto 30 minutes per assay Ths feature is ideal for high-throughput screening (HTS) applicatonsin which as many as 100,000 assays maybe run daily ‘on robotic equipment, Because incubation periods are often a limiting, factor the development of rapid automated assays, SuperSignal ELISA Fomta Substrates the logical choice for automated HTS applicatons. Highlights: + Immediate light generation —intonse signal generated Jimmediately at both room temperature and 37°C + Improved low-end inearty ~easy detection of low quantities often with ih signa rats andow-ond inesty ‘of dase response curves, + High sensitivity ~fermtogram-level detection of target proteins + Reduction in assay time — high sensitivity allows for reduction inincubation steps + Stability storage fr six momths atroom temperature or a minimum ‘of T2months at4°C with a sic-hour werking slution stability + Emits light at 425nm, ge Incubation tine rquved to reach maximum signal Supctgnal LISA Fete Macimom Sst Sabla opines aroonienperate| jownload product instructions, visit www thermoscientiic Nery ae Femtogram detection of tage protein and dose respanse curve generated fom an IL-7 ELSA lusts the exceptional low-end inatty achieved with Thetrno Scene SuperSgnal ELISA Fomto, Subsate andthe incredible sonstvyataabl. ELISA, which was downto Wai. The Rvalue ofthe curve was 120 far signal generated atlers than ‘oat of 2 1200 soo Neen OH ps si ‘Signal intonsity and kinetics comparison ‘The dase response cuve using There Se Maximum Senstivay Suber nan L-2 ELISA was compared to curves ‘generates vith adioeetane-based substrate, another luminob based substat {n acridarbasee system and TMB, SuperSignal EUSA Ferte Maseru Senstniy Subseat demonstrated inedine ght generation with maxenun eakintensty and high RLU values ao hertete ate Aaeanees Bane EB ela J Cn ves M186 167. rly, NAS. aps J. 2001. Phares Ep. Tae me, 47 ap anaCarlissen, CX. 202m Ime 726, etd Chm a tz 182 Pkg, Size 7075 SuperSignal EUSA Femta Maximum ont ‘Senstivty Substrate Inde: Lireralhaner sont. SoneQuantaBlu Fluorogenic Peroxidase Substrates The ideal fluorescent substrates for use with peroxidase enzymes. Avariety of substrates are avalatle for detecting peroxdase actvtyin ELISA-based assays. Colorimetric substrates (e.g, ‘TMB, OPO and ABTS) have been used widely for yeas. Each ofthese substrates varies realy with respectto its performance characteristics such as detection senstiviy, ‘working range and attainable signalnose ratios, Substrate flexibility is also a key ssue that affects ‘an assay. Stabilty, development time requirements and the capacity to perform stopped and/or kinetc assays vary significant among these substrates. Ideal, a substrate is stable, i very sensitive, has high attainable signalose ratios, possesses a broad dynamic range, and allows the user to perform stopped, nonstopped and kinetic assays. ‘Thermo Scientific QuantaBlu Fluorogenic Peroxidase Substrate ‘meets allthe requirements of an ideal substrate for use in peroxidase detection ‘QuantaBlu Substrate goneratesa blue fluorescent product upon ‘waction with peroxidase that does not photobleach. uorametrie- based detection overcomes the iitations of colorimetric substrate dotection, which does notallow for quantitation of greatr than four optical density units. QuantaBlu Substrate alos fr stopped, ‘nonstopped and kinetic assays to be performed, Incubation times for ‘topped and nonstopped assays canbe varied between 1 to 90 minutes at cither room temperature or 37°C. QuartaBlu Substrate exhibits a fat baseline in assays, which faciltates low-level detection sensitivity and allows for high signal noise ratios (se figure top right ‘QuantaBlu Fluorogenic Peroxidase Substrate enables rapid detection cof peroxidase at vey low concentrations. Peroxidase is detected at 0-10pg per wel from 15t0 65 minutes ofsubsizate incubation time. ‘Accyele (15 minate incubation) as litle as 25pg of peroxidase could be detected, while at cycle 6 (55 minute incubation} 0.625pg of peroxidase could be detected (see figure, lower right Highlights: + More sonstve than TMB, OP or ABTS substrates + Flexible topped, nonstopped or kinetic assays possible + Large dynamic range (tog peroxidase cancertration range) + Large Stokos' shit, xctatior/emission maxima of 325/420, range of 215-345/370-460 + Does not photobleach aeons ean Hea 20) re Nel Ace. Se 7, 6848, fi i im at in Py ang oll Py 9, Lo ss Pree a, 68 Seve MD, tal 68, Peies 22) Te SIN Ratio Biya RP oo —o— on ‘Comparison of Thermo Scientific QuantaBlu Substrate to other substrates (QuanaBly Substrate ana the colormete subs ates were ieubated for {Wrinutes araom temperature, flowed by ation of a stop alin, ‘QuartaBla Substrate produced te greatest sinalnoise (INI ati and tes the loves detection it 0 2 SNRs » 1 2 + . + v0 Bloay eed HRP ppl tection with Thermo Scent QuantaBla Fi Peroxidase Substrate Bound btnyleted peroxicase was detected st ‘-palel The assay wos perforin nonstopped mode sing 31-minute insrument cycle time between Citron Product Descriptio Pig. Size 15168 GuantaBluFuorogenic Peroxidase Subsite Kt Sib Bont Feros Soliton Tank a Bak W562 GuantaBlu NS Substrate Ke and kinte assays) ot ont Suara Sable Prd Sion ok To order, call 00174-5728 o 15-56-0747, Outside the United Stats, contact your acl branch ofce or dnb rmQuantaRed Chemifluorescent HRP Substrate Get the ultimate in sensitivity for ELISA-based applications. ‘Thermo Scientific QuantaRed Enhanced Chemifluorescent HRP Substrate enables low picogram levels of detection in the ELISA format. The QuantaRed Substrate uses a proprietary technology to increase fuorescent yield and sensiivity ofthe wel-documented ADHP (10-Acety/-3,7-itydroxyphenoxazine) chemifluorescence reaction, ADHP is a non-fluorescent compound that reacts with horseradish peroxidase (HAP) to produce resorufin, a soluble, highly fuoroscont reaction praduet with oxctation/amission ‘maxima of 670/685nm (Figure 11). The long wavelength emission ofthe resorufin minimizes interference from the low wavelength (blue and green} autofluorescence, making it an ideal substrate for quantitating target molecules in biological samples. In addition, the reaction product of QuantaRed Substrate is colorimetric as well as being fluorescent and can be read spectrophotomevricaly at Sm (Figure 11), Wavelength a Faure Namatzedasepon cl nd nisin seca (the ont HRP Subst, Specially formulated, QuantaRed incorporates chemical enhancers for maximum sensitivity and superior low end linearity (Figure 11). With high quantum yield and large extinetion cootficiont, QuantaRed Enhanced HRP Substrate has sensitivity comparable to enhanced chemiluminescence and is more sensitive than the Amplex Red substrates forthe detection of HRP in ELISA applications. jownload product instructions, visit www thermoscientiic eceninat Hames 7 (pnt) Figure 12 Thermo Scienife QuantaRed Enhanced Chemiuorescent HRP Substrate enables law picogram eget detection. The detecton lino the ‘assay ws 1p (1619 [Panel Bl Als determined by near regression, Pli-stimulateg PBMCs contained T8Sgim hua I-17 andthe urstimuatod samples contaned< 16pgim Highlights: + Sensitive -low picogram to femtogram detection ion 3 sw: i ne ix : » rl Over Tine with tp Soe Figute 1, Fluorescent signal ie stabe for several hours, Four hours ster a ng the Guantaod Stop Solon, the fluorescent sgral was within 3% of he intial irtensty Aer 20 hours, the eacton product maintained neatly 27% of is eign orescence Product Description Pg. Size 15159 Quantaed Enhanced Chenifurescent Kit HRP Substrate “ule rape pra 0x Se Aza {anti ROMP concen int Emancer Slaton Sn Sle Pern Sain Son Sp auton fonABTS: opp ABTS (2.2"azinobis 2-ethylbencothiazoine--sulfnic ac) (OPD yields a water-soluble yellow-orange product when reacted ‘manium satis a water-soluble HRP substrate that yields a gr with peroxidase wit an absorbance maximum of 482. OPD can product upon reaction with peroxidase. The green product has two sy be dissolved in a substate butter such as Therma Scientific ‘major absorbance peaks, 40Snm and 650nm, ABTS i ess sensitive Stable Peroxide Substrate Butfer (Product #24062, than OPD and TMB in ELISA applications tis less readily oxidized, and'ts color developments stower {approximately 0 minutes) This ee ‘may be advantageous'f unacceptable background results from the ‘use ofthe OPD or TMB substrates from higher sensitiv Product Descrnti Py Sice Highlights: 005 __0PD ia pow + One component 00s OPO Ta So ables (g/l) + Ready-to-use + Excellent choice when maximum sensitivities are nat required + Slow reaction that can be easily monitored witha kinetic reader alae THO a a J. Bi. Chm 27,1282 Swng Wl SoC, Age ie Mab, 26208, Product Description Hoa ABTS Desripon See Marinam Cote Proc t Pree Sp ABTS Tow os ‘ain Gen as STS Toke Tow we im Gen 3 OPO Ta OD ca sal aiaahdin a cae ber cy Ne ‘sem Yet ot ‘sn ar Sable Prxde Sbeate Sater 08 Froese ts O70 Powe a Ne em Yet 0% Wa Suber ke ie est | ev tigh Wo ‘sm Yet 7 enya “ets sean, {Ne DM DISD reaper Peres Sup Oe component Vevigh We ‘sm Vote eH tie hevgytue “Hibet ses SHSONE cr MSO nthe oper Pore 3a cy Ne os Yolen en HSDNE ors nthe regent eee Sip One campenent Wein So Yelow ee Seer Sheva To order, call 00174-5728 o 15-56-0747, Outside the United Stats, contact your acl branch ofce or dnb Ls |x MB The ideal fluorescent substrates for use with peroxidase enzymes. TMBis a chromogen that vielés a bue coor measurable st TOnm or Pam) when tied wth hydrogen peroxide catalzod by HRP ‘Tho colorthen changes to yellow (measured at 0hm) Upon addition a suture or phosphoric act stop tho reacton, MB is very sensitive and mare quick oxidized than ‘other HRP substrates resuting in faster color development. ‘The Thermo Scientfic 1-Step TMB Substrates are one-component substrates that require no preparation before use. Unke other commercially available substrates, these products contain no DMIF ‘or DMSO. There are three formulation that cffer primarily intheir sensitivities Step Slow TMB is intermediate in senstviy —ideal for kinetic readings. The sensitivity of the Step Turbo TMB compares to thatof OPO used at approximately Imgiml. 1-Step Ukra TMB-ELISA produces the highest signalnoise ratio and sensitvty inthe picogram range, andhas a three-year shelf if. Hu ii es i i a3 saci 25x 0 asx pean tne gan ‘ren momt jownload product instructions, visit www thermoscientiic Abanbance 0m SO saat ses Tharme Seige 1-Step Ultra TMB-ELISA provides more signal than ther TMB substrates, 1s * ‘Thame coe up ira TM EUSA produces ihe sia nie (SM) ratios than ether TMB substrates andithas a hree-yoar shel Ie eeenes ong, Po 0, ied 285-18, up. AS, oa ie Ais As 1 Su Stal (ae ool her 28, 1812112 Tek Ware Zale Mt Pra et 8218, Thies 26 eta 16 Atal Sacer 5, 8. Weimer 8, Agp En Merb 200207 WU SSE and ong, St Aa Eur Mab TNT, dering Information Product Description Pig. Size ona 20m yo 250m Moan Step Utre TMB-ELISA 20 maar Sa Ki Inelies abet M5) Bink ional 2mAlkaline Phosphatase Substrates PNPP Alkaline phosphatase (API, a 140kDa protein thatis generally ‘solated from calf intestine, catalyzes the hydrolysis of phosphate ‘groups from a substrate molecule, resulting ina colored or fluorescent product or the release of ight as a byproduct. AP has optimal enzymatic activity ata basic pH (pH 8-10) and can be Inhibited by cyanides, arsenate, inorganic phosphate and divalent ‘ation chelators, such as EDTA. As a label or ELISA, AP offers a distinct advantage ovor other enzymes. Because its reaction rate remains linear, detection sensitivity can be improved by simply allowing a reaction to proceed longer. Because ofits large size, [AP is sensitive to freeze/thaw cycles and should be aliquotted and stored at 4°. The most commen ELISA substrate for alkaline phosphatase is the ehromagen p-nitrophenyl phosphate (PNPPI, which is ‘available in several formats. PNP yields a yellow reaction product that is water-soluble and absorbs light at 405m. The Thermo Scientific Pierce 1-Step PNPP Substrate (Product # 37621) offers the convenience of a ready-to-use reagent with similar sensitivity tothe two-component kt. The Phosphatase Substrate Kit (Product #37620) includes PNPP tablets and diethanolamine butfer (5X), PNPP is also available as 259 of powder (Product # ‘34045 or in tablet form (Product #34047). Diethanolamine ‘Substrate Buffer (Product #34064) isa convenient, ready-to-use formulation sold as a 5X concentrate with an optimal and consistent pH, Soluble ELISA substrates for AP are summarized in Table 6, on page 58. Theme Seige Pierce Alkaline Phosphatace Substrates, Kinte Mrssarement Absorbance Posie" Manian” Cole Pruett ve ‘ein Yeon aan ve ‘inn Wlio—a8 1 ‘emer tF Detection of alkaline phosphatase in ELISA applications. NPP (p-nitrophenyl phosphate, disodium salts a widely used substrate for detecting alkaline phosphatase in ELISA applications” ‘When alkaline phosphatase and PNPP are reacted, a yellow water soluble reaction produetis formed, Ths product absorbs light at 405nm. We offer PNPP in four formats. PNPP is avaiable ether as a crystaline powder or mg tablets. Also available i tho Therma Scientfic Phosphatase Substrate Kit which contains PNPP tablets and a Diethanolamine Buffer to yield more than one ter of substrate ‘The Diethanolamine Substrate Butter (Product #34064) is also previded individually as @5X concentrate, Ourformulaton has an ‘optimal and consistent pl and itis stable even at a 1X concentration, Unt recendy, tho method for preparing PNPP solutions required dissolving the powder or tablets in bufer and then diluting the solution ‘othe desired concentration, Substrate instability made tnecessary to prepare tana daily basis, The 1-Step PNPP circumvents this ‘time-consuming and variablesntroducing stop by providing a single component PNPP substrate. This substrate is stable for 12months at 2-8°C and can be stopped with conventional methods Highlight F al 97, Boch Bape. Acta, 176-18 Bove Baal 02 Pre Nat Ae $098,312.87 Senlsir aa a 0a Crm Product Descriptio Pig. Size Hs PAP ‘a ponter WAT PNPP Tobie “stakes [rates ‘RD Phosphate Substrate Kit Ke 2800 osc asi ‘ORE Pierce TStep PNP oem To order, call 00174-5728 o 15-56-0747, Outside the United Stats, contact your acl branch ofce or dnb myONPG Colorimetric {-Galactosidase Soluble Substrate For detection of-Galactosidaso label in ELISA applications. When using f-Galactosidase as the label for proteins in ELISA studies, a wide varity of substrates are available, including o-ntropheny-f-D-galactopyranoside (ONPGI, naphtnol-AS-Bl {-D-galactopyranosie (Nap-Gal) and 4-MethyLumbelifery--D- galactopyrenoside (Mum-Gall However, itis important to choose a substrate with adequate solubility, trat uses readily available {equipment and that give a signficantreading over the background ONPG isa superior f-Galactosidase substrate option” The product formed is completely soluble and hes a high extinction coefficient at 45nm. The substrate yields a yellow productthat is easily dotoctabl in the visual range aftor stopping the reaction with 1M sodium carbonate Highlights: + Forms a completly soluble product when reacted with B-aalactosidase + High extinction cosficent at 45am + Yields yellow product easy detectable inthe visualrange Characteristics of Thermo Scientific Pierce ONPG Substrate, inate NMessronent Ababanee Desrpion _Senitniy Paste” Maxinam Col Prodet ors Hah Yes ‘io ‘ellen 38 eters Merron GR, e965 Bat hom 24, 24824 Ordering Information Product # Descriptio ‘ass _oNPe Pkg. Size Saponier jownload product instructions, visit www thermoscientiic Substrate Buffers ‘Substrate butfors fr alkaline phosphatase and horseradish peroxidase. Butfered stable peroxide is used by researchers who prefer tomake their own HRP-ELISA substrates This is done easily by adding a chromogen of choice tothe buffer. Thermo Scientific Stable Peroxide Substrate Butfer has along shel Ife {12 months ater receipt) and is provided as a 10X concentrate. total volume ‘of 1000mL ean be made from one 100ml bottle of concentrate, If You use TOmL of substrate per plate, you can make substrate for 100 plates from only ana bottle af aur Stable Peroxide Substrata Butter. Our Diethanalamine Substrate Buffer is used with soluble alkaline phosphatase substrates such as PNPP. Our formulation is convenient, ready-to-use and reduces the possibilty of assay-to- assay variability Diethanolamine Substrate Buffers sold as ‘5X concentrate with an optimal, consistent pH. Itis stable even ata IX concentration, Highlights: Ready-to-use ‘Minimize assay-to-assay variability aes teu eal) Hitechon, Cuoco 88-8 Nesewinet det 0 Neos. a E08, Ped tal tt amid W818 tosobles (sngrable 720 Phosphate Substrate Kit Ki Senet tat reat Sep PAP