Histogene immuno_kits_brochure_c.pmd

HistoGeneTM LCM Immunofluorescence Kit
Preserve RNA Integrity during Immunofluorescence Processing
Simplify Fluorescent Target
Cell Labeling

Identify target cells for Laser CaptureMicrodissection (LCM) and gene expressionanalysis using antigen markers to highlightspecific surface or intracellular proteins. TheHistoGene LCM ImmunofluorescenceStaining Kit is the only Kit specifically designedfor retrieval of high-quality RNA fromimmunofluorescently stained frozen tissuesections. The Staining Kit provides reagents andslides for convenient and reliable immuno-fluorescent staining. The Kit provides aprotocol streamlined and optimized to maintainRNA quality and materials sufficient forprocessing 32 slides.
Chill Samples
Get Brilliant High-contrast
for Intact RNA
Label Intensity
The HistoGene LCM Immunofluorescence Kitemploys a biotin-avidin system with Cy3 whichleads to exceptionally good staining intensity andspecificity. The Kit is used with a primarybiotinylated monoclonal antibody to an antigen ofchoice provided by the user and Cy3-conjugated Laser Capture Microdissection (LCM) of human prostate ductal
epithelium prepared with the HistoGene LCM Immuno-
streptavidin is included in the Kit.
fluorescence Kits. Panel A. Human prostate stained with an anti-
cytokeratin antibody, before LCM. Panel B. Image of the captured
cytokeratin positive cells on the cap after LCM.
Use A Quick 15-minute
Process to Preserve RNA

To increase RNA yield and ensure its quality, the HistoGene Cold Block keeps up to four generally take over 90 minutes to process tissue section slides and several tubes containing samples, compromising the RNA integrity with buffer and antibody solutions chilled during conditions sub-optimal for single-stranded processing. The block is designed for use with Identify target cells using the HistoGene LCM Immuno-
the CoolSafe triple density polysterene cooler protocol takes less than 15 minutes (only 5 fluorescence Staining Kit. Panel A. Human foreskin stained with
(Cat. # CSF-Box) and –10ºC CoolBrick (Cat. # anti-CD1a antibody. Panel B. Human jejunum stained with anti-Pan-
cytokeratin antibody. Panel C. Mouse lacrimal gland inflammatory
performed using Arcturus’ proprietary staining infiltrate stained with anti-CD4 antibody. Panel D. Mouse small
buffer, which minimizes RNA degradation.
intestine stained with anti- MAdCAM-1 antibody.
Electrophoresis and RT-PCR of RNA isolatedafter using the HistoGene Immuno-fluorescence Kit confirm that high-qualityRNA is retrieved from microdissected samples.
(Data on back page.) Comparison of RNA quality before and
Obtain Intact RNA from
after processing using the HistoGene
LCM Immunofluorescence Kit. RNA was
Many Tissue Types
Arcturus scientists have examined RNA integrity (Lane 1), and from frozen section of small intestine stained using the HistoGene LCM after using the HistoGene Immunofluorescence Kit on many tissue types with several antibodies.
vascular expression of MAdCAM-1 (Lane 2).
All tissue tested using the HistoGene Immuno- Electrophoretic analysis of total cellular fluorescence Kits have yielded high-quality RNA.
Detection of low- and high-abundance, full-length RNA in
HistoGene LCM immunofluorescent stained samples. RT-PCR was
performed on RNA from 500–1000 cells captured from different Validated Tissue-Antibody Sets
tissues. Equal quantities of cDNA were analyzed with primers detecting 3’ and 5 ‘ ends of low-abundance ADP ribosylation factor Antibody
1 ( ARF-F1), (3’; 239 bp, 5’; 336 bp) and 5’ end clathrin gene which Use Microgenomics Products
has a transcript length over 6 kb (570 bp) (primer sets from KPL, Inc.). M: molecular-weight markers. Lane 1: human breast tissue, ARF- for Microarray Analysis
F1, 5’. Lane 2: mouse brain, ARF-F1, 5’. Lane 3: mouse spleen, ARF-F1, 3’. Lane 4: mouse lacrimal gland, ARF-F1 3’. Lane 5: mouse lacrimal gland, clathrin. Lane 6: human prostate, clathrin. Lane 7: negative The HistoGene LCM Immunofluorescence Kit is RT control, human prostate, clathrin.
Microgenomics–products designed to seamlessly work together to produce high-quality expression microarray data from pure cell populations.
Use HistoGene-stained samples with thePixCell® IIe LCM System or the AutoPix®Automated LCM System to capture pure cellpopulations of fluorescently labeled target cells.
Ordering Information
Expression microarray of LCM samples prepared using Arcturus’
Catalog #
numbers of cells with the PicoPure RNA HistoGene LCM Immunofluorescence, PicoPure RNA Isolation,
Isolation Kit. Next, amplify nanogram quantities and RiboAmp® RNA Amplification Kits. Human breast tumor
Staining Kit, reagents for staining 32 slides of RNA to micrograms using the RiboAmp® RNA tissue sections were stained for cytokeratin using the HistoGeneAmplification Kit, which provides amplified LCM Immunofluorescence Kit and 1000- positively stained cells were microdissected. RNA was isolated using the PicoPure RNA Isolation Kit and amplified with the RiboAmp RNA Amplification Kit.
Amplified RNA was converted to Cy5 labeled cDNA and hybridized Related Products
to a 24,000 element human cDNA array and scanned.
Catalog #
HistoGene LCM Immunofluorescence
Kit Components
· HistoGene Buffer A (4 ml)· HistoGene Buffer B (60 ml) · HistoGene LCM Immunofluorescence Staining RiboAmp® HS RNA Amplification Kit,enough for 5 two-round amplifications PicoPure RNA Isolation Kit, enough for40 isolations RiboAmp, PicoPure, HistoGene, PixCell and CapSure are trademarks owned by Arcturus. All other trademarks shown are the property of their respective owners. Polymerase Chain Reaction (PCR) is a patented process owned by Hoffman-La Arcturus Bioscience, Inc.
400 Logue Avenue

Source: http://parallabs.vss.ru/pdf/HistoGene_Immuno_Kits_Brochure.pdf

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