IHC Polymer
Presented for: BioGenex Website
Presented by: BioGenex Tech Support
Date: 2016
Immunohistochemistry (IHC) Quick Guide#
Super SensitiveTM Polymer-HRP kits.
Baking
Overnight,50-60oC
then
15-30min,60-80oC
Beforestaining
An-gen
Retrieval
DeWax
EZ-DeWax3x3min,RT
20min,RT
Peroxide
Block
For95%ofBioGenexan-bodies
UseEZ-AR-2Elegance-20min,95oC
SeeCatalogforan-body-specific
recommenda-on
5-10min,RT
PowerBlock
0-10min,RT
Super
Enhancer##
Primary
An-body
Polymer
HRP
For>90%ofBioGenex
30min,RT
an-bodies:30min@RT
or
20min,37oC
SeeDatasheetfor
an-body-specific
Incuba-oncondi-ons
DAB
5-10min,RT
Hematoxylin
1-3min,RT
Moun-ng
SuperMount®Medium
Or
Xmount(forAutoma-on)
Ki-67staininginTonsil/ssue
#FormanualusewithFFPE-ssueslides.Washingstepsomibedforsimplicity-forfullprotocolseePoly-HRPkitdatasheet.Op-miza-on
mayberequiredduetovaria-onin-ssues/processing.
##NotRequiredwhenusingOneStepPoly-HRPDetec-onSystem
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Immunohistochemistry (IHC) Quick Guide
Super SensitiveTM Polymer-HRP kits.
1. Baking – Hea-ng the -ssue slide helps remove humidity
and promote abachment of -ssue to slide. It also helps wax
removal in the following De-Wax step. Depending on -ssue
type and thickness, incuba-on temperature and -me may
vary.
2. DeWax – BioGenex EZ-DeWax Solu-on (HK585-5K &
HK584-5K) is a “one-step” product that simultaneously
enablestheremovalofparaffinandallowsrehydra-onofthe
-ssuewithasinglexylene-free,non-toxicreagent.
3. An/gen-Retrieval BioGenex is the inventor of An-gen
Retrieval enabling technology. An-gen-retrieval step breaks
the formalin induced cross-linking bonds between epitopes
and unrelated proteins, thereby allowing beber penetra-on
of an-body and accessibility of epitopes. BioGenex solu-ons
facilitatetheproduc-onofhighlyreproducibleandsuperior
qualitystainsinaconsiderablyshortperiodof-mewithoutcompromisingthemorphologyandan-genicityofthe-ssue.TheEZ-AR2EleganceSolu-onadvancedformula-on(HX032-YCD&HK547-XAK)hasuniquesuperhea-ngproper-esandwasvalidatedtogivethe
beststainingwith95%ofBioGenexAn-bodies(seedatasheetforAb-specificrecommenda-on).
4.PeroxideBlock(HK111-50K)–Hydrogenperoxideisusedtoinac-vateendogenousperoxidasewhenHorseradishPeroxidaseisused
forstaining.Thiseliminatesbackground,especiallyin-ssuessuchasredbloodcells,kidney,andliver.
5. Power Block (HK083-50K) – This reagent contains buffer, casein and preserva-ve and reduces background staining due to nonspecificbindingofan-bodies.Itisapotentblockerandshouldnotremainonthe-ssuemorethantenminutes.
6.PrimaryAn/body-BioGenexoffers>400an-bodiesforIHCstaining.An-bodiesareavailableinready-to-use(RTU)orconcentrated
formats and are for manual or automated staining. BioGenex an-bodies are op-mized to provide maximum signal with minimum
background.
7.Polymer-HRP–Novel,readytouse,universalan-bodydetec-onsystemusinganon-bio-npolymerictechnology(seefigure).This
reagentcontainsamixtureofgoatan--rabbitandan--mousesecondaryan-bodies
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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 enhance
binding.Asthesystemisnotbasedonthebio-n-avidinsystem,problems
associated with endogenous bio-n are completely eliminated. The
technology allows excellent cell penetra-on ability compared with other
polymer-HRPs.
8.DAB(HK542-XAKE)–DAB(3,3'-diaminobenzidinetetrahydrochloride)is
achromogenicperoxidasesubstrate.HRPcatalyzestheoxida-onofDAB
toformabrownprecipitatewhichstainsthe-ssueincloseproximityto
theHRPconjugatedan-body.DABisinsolubleinalcohol,xyleneorwater
and therefore is compa-ble with most clearing agents and moun-ng
media.
A.An--MUC5AC
B.An--CD3(T-Cell)
C.An--Glypican-3
9.Hematoxylin(HK100-5K,HK100-9K)–(BioGenexMayer’shematoxylinisabluenuclearcounterstainthatmakesthestained-ssues
moreeasilyvisible.Itisalcoholandxyleneinsolubleandthereforecompa-blewithmostclearingagentsandmoun-ngmedia.
10.Moun/ng-Moun-ngsealsthebiologicalspecimensandisanessen-alstepbeforemicroscopicevalua-on.BioGenexSuperMount®
PermanentMoun-ngMedium(HK079-5K,HK079-7K)isapolymerbasedaqueousmoun-ngmediathatdoesnotrequiretheuseofa
coverslip.XMountTMMoun-ngMedium(HX035-YCD,HX035-10X)isapermanentmoun-ngmediumthathasbeenop-mizedforuse
withBioGenexXmatrxautoma-onplasorm.BioGenexalsooffersAqueousMoun-ngMedium(HK099-5K)-aglycerol-basedmoun-ng
mediumthatrequiretheuseofacoverslip.Itisintendedforshort-termspecimenstorageandiscompa-blewithmostchromogens
andcounterstains.
11.Washingsteps–Typically2-3washesbetweeneachsteparerequired.BioGenexSuperSensi-veTMWashBuffer(HK583-5K)isa
phosphate buffered saline (PH 7.4) with surfactant and is the best choice in order to ensure op-mal staining. The buffer promotes
evenspreadingofan-bodiesandotherreagentstoavoidinconsistentresults.
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BioGenex Immunohistochemistry (IHC) Quick Guide
Super SensitiveTM Link-Label kits
Formanualuse(dropbobles)
Fori6000Automa-on(Barcodelabeled)
ForXmatrxAutoma-on(Elite-RFIDtaggedvials)
Detec-onSystems-SuperSensi-vePolymerHRP(Formouse&rabbitan-bodies)
SKU
QD400-60KE
QD420-YIKE
Size
60
test
500
test
Super
PolymerDABbuffer
enhancer HRP
DAB
Peroxide Power
Chromo. block
block
Hematox.
EZ-AREleg.
Nega/vectrlIgG
1,2&3Sol.
6ml
6ml
10ml
2ml
6ml
6ml
6ml
-
3ml-Rabbit,3ml-Mouse
50ml
50ml
50ml
5ml
-
-
-
-
-
QD430-XAKE
1000test 100ml
100ml
100ml
10ml
-
-
-
-
-
QD440-XAKE
1000test 100ml
100ml
-
-
-
-
-
-
-
200test
20ml
20ml
5x10ml
4ml
20ml
20ml
20ml
-
-
200test
15ml
15ml
5x11ml
4ml
4x16ml 21ml
21ml
16ml
-
3x16ml
3x16ml
-
QD410-YAXE
i6000
QD550-YCXE
Xmatrx-Infinity
QD550-YCDE
Xmatrx-Elite
IHCDetec-onSystems-Links/Labelsitems-Manual
4x11ml+5RFID
200test 16ml
16ml
7ml
3x16ml 3x16ml
vials
Detec-onSystems-SuperSensi-veOne-StepPolymer-HRP(Formouse&rabbitan-bodies)
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
-
-
-
-
200test
16ml
4x11ml
4ml
3x16ml
3x16ml
3x16ml
-
200test
16ml
4x11ml+5RFIDvials 7ml
3x16ml
3x16ml
3x16ml
3x16ml
QD610-YAXE
i6000
QD610-YADE
Xmatrx-Elite
ThankYou
Pleasevisitwww.biogenex.comformoredetailsonourproductporsolio
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