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IHC Polymer

BioGenex Tech Support

2016

BioGenex Website

Baking DeWax An-genRetrieval

PeroxideBlock PowerBlock

Immunohistochemistry (IHC) Quick Guide#

Super SensitiveTM Polymer-HRP kits.

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PrimaryAn-body

PolymerHRP DAB Hematoxylin Moun-ng

Overnight,50-60oCthen

15-30min,60-80oCBeforestaining

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

SeeCatalogforan-body-specific

recommenda-on

5-10min,RT 0-10min,RT

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

SeeDatasheetforan-body-specific

Incuba-oncondi-ons

30min,RTor

20min,37oC

5-10min,RT 1-3min,RT SuperMount®MediumOr

Xmount(forAutoma-on)

SuperEnhancer##

20min,RT

Ki-67staininginTonsil/ssue

#FormanualusewithFFPE-ssueslides.Washingstepsomibedforsimplicity-forfullprotocolseePoly-HRPkitdatasheet.Op-miza-onmayberequiredduetovaria-onin-ssues/processing.

##NotRequiredwhenusingOneStepPoly-HRPDetec-onSystem

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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).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.

1. Baking – Hea-ng the -ssue slide helps remove humidityandpromoteabachmentof-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. An/gen-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 beber penetra-onofan-bodyandaccessibilityofepitopes.BioGenex solu-onsfacilitatetheproduc-onofhighlyreproducibleandsuperior

6.PrimaryAn/body-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.Polymer-HRP–Novel,readytouse,universalan-bodydetec-onsystemusinganon-bio-npolymerictechnology(seefigure).Thisreagentcontainsamixtureofgoatan--rabbitandan--mousesecondaryan-bodies

Immunohistochemistry (IHC) Quick Guide Super SensitiveTM Polymer-HRP kits.

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

7.(cont.)conjugatedtoapolymerizedhorseradishperoxidase(Poly-HRP).The 2-step kit requires pre-incuba-on with Super Enhancer to enhancebinding.Asthesystemisnotbasedonthebio-n-avidinsystem,problemsassociated with endogenous bio-n are completely eliminated. Thetechnologyallowsexcellent cellpenetra-onability comparedwithotherpolymer-HRPs.

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.Moun/ng-Moun-ngsealsthebiologicalspecimensandisanessen-alstepbeforemicroscopicevalua-on.BioGenexSuperMount®PermanentMoun-ngMedium(HK079-5K,HK079-7K)isapolymerbasedaqueousmoun-ngmediathatdoesnotrequiretheuseofacoverslip.XMountTMMoun-ngMedium(HX035-YCD,HX035-10X)isapermanentmoun-ngmediumthathasbeenop-mizedforusewithBioGenexXmatrxautoma-onplasorm.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--MUC5ACB.An--CD3(T-Cell)C.An--Glypican-3

Formanualuse(dropbobles) Fori6000Automa-on(Barcodelabeled) ForXmatrxAutoma-on(Elite-RFIDtaggedvials)

Detec-onSystems-SuperSensi-vePolymerHRP(Formouse&rabbitan-bodies)

IHCDetec-onSystems-Links/Labelsitems-Manual

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

SKU Size Superenhancer

Polymer-HRP DABbuffer DAB

Chromo.Peroxideblock

Powerblock Hematox. EZ-AREleg.1,2&3Sol. Nega/vectrlIgG

QD400-60KE 60test 6ml 6ml 10ml 2ml 6ml 6ml 6ml - 3ml-Rabbit,3ml-Mouse

QD420-YIKE 500test 50ml 50ml 50ml 5ml - - - - -

QD430-XAKE 1000test 100ml 100ml 100ml 10ml - - - - -

QD440-XAKE 1000test 100ml 100ml - - - - - - -

QD410-YAXEi6000 200test 20ml 20ml 5x10ml 4ml 20ml 20ml 20ml - -

QD550-YCXEXmatrx-Infinity 200test 15ml 15ml 5x11ml 4ml 4x16ml 21ml 21ml 16ml -

QD550-YCDEXmatrx-Elite 200test 16ml 16ml 4x11ml+5RFID

vials 7ml 3x16ml 3x16ml 3x16ml 3x16ml -

SKU Size Polymer-HRP DABbuffer DABChromo. Peroxideblock Powerblock Hematox EZ-AREleg.1,2&3Sol.

QD620-YIKE 500test 50ml 50ml 5ml - - - -

QD630-XAKE 1000test 100ml 100ml 10ml - - - -

QD610-YAXEi6000 200test 16ml 4x11ml 4ml 3x16ml 3x16ml 3x16ml -

QD610-YADEXmatrx-Elite 200test 16ml 4x11ml+5RFIDvials 7ml 3x16ml 3x16ml 3x16ml 3x16ml

Detec-onSystems-SuperSensi-veOne-StepPolymer-HRP(Formouse&rabbitan-bodies)

ThankYouPleasevisitwww.biogenex.comformoredetailsonourproductporsolio

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