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IHC Link-Label

BioGenex Technical Support

2016

BioGenex Website

BioGenex Immunohistochemistry (IHC) Quick Guide# Super SensitiveTM Link-Label kits

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Baking DeWax An-genRetrieval PeroxideBlock PowerBlock

PrimaryAn-body Link Label DAB Hematoxylin Moun-ng

For>90%ofBioGenexan-bodies:30min@RT

SeeDatasheetforan-body-specific

Incuba-oncondi-ons

10-20min,RT

5-10min,RT 1-3min,RT20-30min,RT

ExampleofCytokera0n18StaininginFFPE0ssue

#ForusewithFFPE-ssueslides.WashingstepsomiXedforsimplicity-forfullprotocolseeLink-Labelkitdatasheet.Op-miza-onmayberequiredduetovaria-onin-ssues/processing.

Overnight,50oCthen

15-30min,60-80oCBeforestaining

EZ-DeWax3x3min,RT For95%ofBioGenexan-bodiesUseEZ-AR-2Elegance-20min,95oC

SeeCatalogforan-body-specific

recommenda-on

5-10min,RT 5-10min,RT

SuperMount®MediumOr

Xmount(forAutoma-on)

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qualitystainsinaconsiderablyshortperiodof-mewithoutcompromisingthemorphologyandan-genicityofthe-ssue.TheEZ-AR2-EleganceSolu-onadvancedformula-on(HX032-YCD&HK547-XAK)hasuniquesuperhea-ngproper-esandwasvalidatedtogivethebeststainingwith95%ofBioGenexAn-bodies(seedatasheetforAb-specificrecommenda-on).

1. Baking – Hea-ng the -ssue slide helps remove humidityandpromoteaXachmentof-ssue to slide. Italsohelpswaxremoval in the following De-Wax step. Depending on -ssuetype and thickness, incuba-on temperature and -me mayvary.

2. DeWax – BioGenex EZ-DeWax Solu-on (HK585-5K &HK584-5K) is a “one-step” product that simultaneouslyenablestheremovalofparaffinandallowsrehydra-onofthe-ssuewithasinglexylene-free,non-toxicreagent.

3. An0gen-Retrieval - BioGenex is the inventor of An-genRetrieval enabling technology. An-gen-retrieval step breaksthe formalin induced cross-linking bonds between epitopesand unrelated proteins, thereby allowing beXer penetra-onofan-bodyandaccessibilityofepitopes.BioGenex solu-onsfacilitatetheproduc-onofhighlyreproducibleandsuperior

BioGenex Immunohistochemistry (IHC) Quick Guide Super SensitiveTM Link-Label kits

4.PeroxideBlock(HK111-50K)–Hydrogenperoxideisusedtoinac-vateendogenousperoxidasewhenHorseradishPeroxidaseisusedforstaining.Thiseliminatesbackground,especiallyin-ssuessuchasredbloodcells,kidney,andliver.

5. PowerBlock (HK083-50K) – This reagent contains buffer, casein andpreserva-ve and reduces background staining due to non-specificbindingofan-bodies.Itisapotentblockerandshouldnotremainonthe-ssuemorethantenminutes.6.PrimaryAn0body-BioGenexoffers>400an-bodiesforIHCstaining.An-bodiesareavailableinready-to-use(RTU)orconcentratedformats and are for manual or automated staining. BioGenex an-bodies are op-mized to providemaximum signal withminimumbackground.

7. Link-Label–Aclassic systembasedon thehighly specificand sensi-ve streptavidin-bio-n interac-on (e.g.QA900-9LE,QP900-9L,QD470-YCXE).

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BioGenex Immunohistochemistry (IHC) Quick Guide

Super SensitiveTM Link-Label kits

7. (cont.) LINK reagent is a mix of an--mouse and an--rabbit IgGsconjugated tomul-plebio-nmolecules. TheLINKan-bodiesdetectandbindtothean-gen-boundprimary-an-body.

LABEL reagent is Streptavidin conjugated with the enzyme Horseradishperoxidase(HRP)(kitswithAlkalinePhosphatase(AP)arealsoavailable).Streptavidin binds extremely strongly and irreversibly to the bio-nresidueson the secondary an-body resul-ng in the addi-onofmul-pleenzymestotheprimaryan-bodycomplex.

8.DAB(HK542-XAKE)–DAB(3,3'-diaminobenzidinetetrahydrochloride)isachromogenicperoxidasesubstrate.HRPcatalyzes theoxida-onofDABto formabrownprecipitatewhichstains the-ssue incloseproximity totheHRPconjugatedan-body.DABisinsolubleinalcohol,xyleneorwaterand therefore is compa-ble with most clearing agents and moun-ngmedia.9.Hematoxylin(HK100-5K,HK100-9K)–(BioGenexMayer’shematoxylinisabluenuclearcounterstainthatmakesthestained-ssuesmoreeasilyvisible.Itisalcoholandxyleneinsolubleandthereforecompa-blewithmostclearingagentsandmoun-ngmedia.

10.Moun0ng-Moun-ngsealsthebiologicalspecimensandisanessen-alstepbeforemicroscopicevalua-on.BioGenexSuperMount®PermanentMoun-ngMedium(HK079-5K,HK079-7K)isapolymerbasedaqueousmoun-ngmediathatdoesnotrequiretheuseofacoverslip.XMountTMMoun-ngMedium(HX035-YCD,HX035-10X)isapermanentmoun-ngmediumthathasbeenop-mizedforusewithBioGenexXmatrxautoma-onplauorm.BioGenexalsooffersAqueousMoun-ngMedium(HK099-5K)-aglycerol-basedmoun-ngmediumthatrequiretheuseofacoverslip.Itisintendedforshort-termspecimenstorageandiscompa-blewithmostchromogensandcounterstains.

11.Washingsteps–Typically2-3washesbetweeneachsteparerequired.BioGenexSuperSensi-veTMWashBuffer(HK583-5K) isaphosphatebufferedsaline (PH7.4)withsurfactantand is thebestchoice inorder toensureop-mal staining.Thebufferpromotesevenspreadingofan-bodiesandotherreagentstoavoidinconsistentresults.

A.An--CK18B.An--GCDFP-15C.An--ErbB-2/Her2

Super Sensitive™ IHC Detection System kits Composition

SKU Size Mul0-Link Label DABbuffer DABChromogen Peroxideblock Powerblock Hematox Nega0vectrlIgG

QA900-9LE 500test 50ml AP50ml - - - - - -

QP900-9LE 500test 50ml HRP50ml - - - - - -

QP300-XAKE 1000 100ml HRP100ml - - - - - -

LP000-ULE 1000+ 5ml(Conc.) HRP5ml(Conc.) - - - - - -

LA000-ULE 1000+ 5ml(Conc.) AP5ml(Conc.) - - - - - -

QD470-YCXE-i6000 200test 20ml HRP20ml 5x20ml 4ml 20ML 20ml 20ml -

Formanualuse(dropboXles) Fori6000Automa-on(Barcodelabeled)

Detec-onSystems-LINKLABEL(Formouse&rabbitan-bodies)

IHCDetec-onSystems-Links/Labelsitems-ManualProduct 5mL(Conc.) 6ml(RTU) 50ml(RTU)

SSAPlabel HK321-UK HK331-5K HK331-9K

SSGoatLink HK327-UG HK337-5G N/A

SSHRPlabel HK320-UK HK330-5K HK330-9K

SSMouseLink HK325-UM HK335-5M HK335-9M

SSMul-Link(ANTI-mouse&rabbit) N/A HK340-5K HK340-9K

SSRabbitLink HK326-UR HK336-5R HK336-9R

SSRatLink N/A HK338-5T N/A

Conc.Mul-Link(ANTI-Mouse,GuineaPig,RatandRabbit) HK268-UK N/A N/A

ThankYouPleasevisitwww.biogenex.comformoredetailsonourproductporuolio

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