Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | None/None |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | RT |
Category | MicroBIOLOGy FluorescenceImaging |
Related | BiochemicalAssays |
1. Platecellsonslidechambersorsterileglasscoverslips,andtreatthecellsasdesired
2. Performformaldehydefixation.Incubatecellswith3.0–4.0%formaldehydeinPBSatroomtemperaturefor10–30minutes.
3. Washthefixedcells2–3timesinPBS.
Optional:Add0.1%TritonX-100inPBSintofixedcellsfor3to5minutestoincreasepermeABIlity.Rinsethecells2–3timesinPBS.
4. Blockwithblockingagentsuchaswith5%BSAinPBSfor30min.
5. Diluteprimaryantibodyindilutionbufferasrecommendedinthespecificproduct’sdatasheet.Overlayenoughdilutedantibodytocovercellsoncoversliporeachchamberofthechamberslides.
6. Wash2-3timeswithPBS.
7. WarmupFluoroQuest™fluorescencesignalenhancingsolutionatroomtemperaturebeforeuse.
8. DilutefluorescentsecondaryantibodyinFluoroQuest™fluorescencesignalenhancingsolutionandthenstaincellsfor1houratroomtemperature.Generalrangeforsecondaryantibodiesisbetween0.1-10µg/mLforIgGconjugatesformostapplications. Keepslipscoveredorinahumidifiedchambertoavoidevaporation.
9. WashthecellsthreetimeswithPBS.
Note:Additionalstainingwithfluorescentnuclearstainsorphalloidinscanbedoneatthisstep.
10. Inverteachcoverslipontoacleanedslidewithmountingmedia,preferablyonewithananti-fadepreservative.Sealedgeswithclearpolishifdesired.
11. Imagingthecellsundermicroscopewithcorrectfilters.Storeslidesinthedarkat4°C.
References&Citations | ![]() PrinterFriendlyVersion |
1. CordesT,VogelsangJ,TinnefeldP.(2009)OnthemechanismofTroloxasantIBLinkingandantibleachingreagent.JAmChemSoc,131,5018.
2. Gallardo-EscarateC,Alvarez-BorregoJ,VonBrandE,DupreE,DelRio-PortillaMA.(2007)RelationshipbetweenDAPI-fluorescencefADIngandnuclearDNAcontent:AnalternativemethodtoDNAquantification?BiolRes,40,29.
3. WidengrenJ,ChmyrovA,EggelingC,LofdahlPA,SeidelCA.(2007)Strategiestoimprovephotostabilitiesinultrasensitivefluorescencespectroscopy.JPhysChemA,111,429.
4. DikiciogluE,MeteogluI,OkyayP,CulhaciN,KacarF.(2003)Thereliabilityoflong-termstorageofdirectimmunofluorescentstainingslidesatroomtemperature.JCutanPathol,30,430.
5. El-RifaiWE,KnuutilaS.(2001)Comparativegenomichybridizationtechnique.MethodsMolMed,50,25.
6. BerriosM,ConlonKA,ColfleshDE.(1999)Antifadingagentsforconfocalfluorescencemicroscopy.MethodsEnzymol,307,55.
7. BrockR,HamelersIH,JovinTM.(1999)ComparisonoffixationprotocolsforadherentculturedcellsappliedtoaGFPfusionproteinoftheepidermalgrowthfactorreceptor.Cytometry,35,353.
8. KolankoCJ,PyleMD,LoatsH,PartonJ,BlakelyWF,NathJ.(1999)Fast-insituhybridizationandimmunoenzymaticcolorpigmentdetectionofmousebonemarrowmicronucleus.BiotechHistochem,74,111.
9. BenchaibM,DelormeR,PluvinageM,BryonPA,SouchierC.(1996)Evaluationoffivegreenfluorescence-emittingstreptavidin-conjugatedFluorochromesforuseinimmunofluorescencemicroscopy.HistochemCellBiol,106,253.
10. SwartzDJ,SantiPA.(1996)ImmunofluorescentartifactsduetothepHofantifadingmountingmedia.Biotechniques,20,398.
AAT Bioquest常用产品报价单
产品名 | 货号 | 公司分类 | 商城分类 | 美金价 |
AAT Bioquest/Fluorescein, disodium salt *CAS 518-47-8*/2/100 mg | 2 | iFluor Rapid Tests | 酸碱缓冲液 | 1088.00 | AAT Bioquest/14-3-3 ζ (Ab-58) Antibody/8B0001/50 ug | 8B0001 | Signaling Intermediate Ab | 重组抗体 | 2828.00 | AAT Bioquest/Opioid Receptor (Phospho-Ser375) Antibody/8A0022/50 ug | 8A0022 | Phospho-Specific Ab | 受体 | 2828.00 | AAT Bioquest/iFluor™ 350 goat anti-mouse IgG (H+L)/16440/200 ug | 16440 | Fluorescent Anti-IgGs | 荧光染料 | 1088.00 | AAT Bioquest/trFluor™ Eu goat anti-mouse IgG (H+L)/16518/100 ug | 16518 | Fluorescent Anti-IgGs | 抗小鼠 | 4278.00 | AAT Bioquest/RPE-streptavidin conjugate/16900/100 ug | 16900 | Streptavidin Conjugates | 其他生物染料 | 1378.00 | AAT Bioquest/GCNT3 Antibody/8C14708/50 ug | 8C14708 | Signaling Intermediate Ab | 功能性抗体 | 2828.00 | AAT Bioquest/Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow*/12518/1 plate | 12518 | Reporter Gene Enzymes | 其它检测试剂盒 | 2103.00 | AAT Bioquest/ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 &mi | 1100 | mFluor™ Dyes and Kits | 标记试剂盒 | 2103.00 | AAT Bioquest/Amplite™ Colorimetric Urea Quantitation Kit *Blue Color*/10058/200 Tests | 10058 | Diagnostic Molecules | 定量试剂盒 | 2828.00 | AAT Bioquest/Screen Quest™ CHO-Gqi Chimera Cell line/38101/Each | 38101 | cAMP GPCR Assays | 常规细胞株 | 0.00 |