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Ausgangstext - Englisch INTENDED USE
PCNA Staining kits are used for the qualitative detection of PCNA protein in formalin-fixed, paraffin-embedded and frozen tissues.
STORAGE AND SHELF LIFE
Store kit at 2-8°C. All performance claims are void after kit expiration date.
REAGENTS SUPPLIED
Reagent 1. One dropper bottle (6 ml) of ready-to-use blocking solution
Reagent 2. One dropper bottle (6 ml) of ready-to-use biotinylated Ms x PCNA primary antibody
Reagent 3. One dropper bottle (6 ml) of ready-to-use streptavidin-peroxidase
Reagent 4A. One dropper bottle (3 ml) of concentrated substrate buffer (20x)
Reagent 4B. One dropper bottle (3 ml) of concentrated chromogen solution, DAB (20x)
Reagent 4C. One dropper bottle (3 ml) of 0.6% hydrogen peroxide (20x)
Reagent 5. One dropper bottle (6 ml) of ready-to-use Hematoxylin
Reagent 6. One dropper bottle (6 ml) of ready-to-use Histomount™
Also contains: 5 Control Slides – 1 stained, 4 unstained
REAGENTS AND MATERIALS REQUIRED BUT NOT PROVIDED
– Phosphate Buffered Saline (PBS)
– Alcohol and xylene
– Distilled or deionized water
– Quenching solution for endogenous peroxidase
– Coverslips
SUGGESTED STAINING PROTOCOL
A. PRELIMINARY PREPARATION
For Paraffin-Embedded Tissues
1. Fix target tissue in 10% NBF (Neutral Buffered Formalin), or in an alcohol-based fixative (such as alcohol or Methacarn). Process tissues for paraffin embedding.
2. Cut tissue 3-4 microns and place on Zymed’s HistoGrip™ (Cat. No. 00-8050) or poly-L-lysine coated slides. Dry slides in a 60°C oven for 1-2 hours.
3. Deparaffinize slides in 2 changes of xylene for 5 minutes each. Rehydrate slides in a series of graded alcohol.
4. (Optional): Block for endogenous peroxidase activity with 3% H2O2 in methanol for 10 minutes. Rinse slides in 3 changes of PBS for 2 minutes each.
5. Slides are now ready for PCNA staining.
6. (Optional): Specific staining of formalin-fixed, paraffin-embedded tissues may be enhanced using a Heat Induced Epitope Retrieval (HIER) method.
7. (Optional): If alcohol-fixed tissues are used, tissue sections can be post-dipped in 10% NBF for 30-60 seconds. Rinse well. This may improve morphology.
For Cultured Cells and Cell Suspensions
1. Fix cells in 70% alcohol or acid-ethanol for 15-20 minutes at 4°C. Acetone or Methacarn fixatives can also be used.
2. If necessary, block for endogenous peroxidase activity with 3% H2O2 in methanol for 10 minutes.
3. Wash in 3 changes of PBS for 2 minutes each.
Übersetzung - Italienisch SCOPI D’UTILIZZO
PCNA I kit di Colorazione sono utilizzati per la rilevazione qualitativa della proteina PCNA in tessuti fissati in formalina, inclusi in paraffina e tessuti congelati.
CONSERVAZIONE E DURATA A MAGAZZINO
Conservare il kit a 2-8°C. Eventuali reclami dopo la data di scadenza indicata sono nulli.
REAGENTI FORNITI
Reagente 1. Una boccetta contagocce (6 ml) di soluzione bloccante pronta all’uso
Reagente 2. Una boccetta contagocce (6 ml) di anticorpo primario biotinilato Ms x PCNA pronto all’uso
Reagente 3. Una boccetta contagocce (6 ml) di streptavidina - perossidasi pronta all’uso
Reagente 4A. Una boccetta contagocce (3 ml) di tampone substrato concentrato (20x)
Reagente 4B. Una boccetta contagocce (3 ml) di soluzione cromogeno concentrato, DAB (20x)
Reagente 4C. Una boccetta contagocce (3 ml) di 0.6% perossido di idrogeno (20x)
Reagente 5. Una boccetta contagocce (6 ml) di Ematossilina pronta all’uso
Reagente 6. Una boccetta contagocce (6 ml) di Histomount™ pronto all’uso
Contiene anche: 5 Vetrini di Controllo – 1 colorato, 4 non colorati
REAGENTI E MATERIALI NECESSARI MA NON FORNITI
– Soluzione fisiologica tamponata al fosfato (PBS)
– Alcool e xilene
– Acqua Distillata oppure deionizzata
– Soluzione Quenching per perossidasi endogena
– Coverslips
PROTOCOLLO DI COLORAZIONE SUGGERITO
A. PREPARAZIONE PRELIMINARE
Per tessuti inclusi in paraffina
1. Fissare il tessuto bersaglio in 10% NBF (Formalina Neutra Tamponata), o in un fissante a base di alcool (come alcool oppure Methacarn). Preparare i tessuti per l’inclusione in paraffina.
2. Tagliare i tessuti 3-4 micron e porli su Zymed’s HistoGrip™ (Cat. No. 00-8050) oppure vetrini rivestiti di poly-L-lysine . Far asciugare i vetrini in forno a 60°C per 1-2 ore.
3. Sparaffinizzare i vetrini in 2 cambi di xilene per 5 minuti ciascuno. Reidratare i vetrini in una scala crescente di alcool .
4. (Facoltativo): Bloccare per perossidasi endogena con 3% H2O2 in metanolo per 10 minuti. Risciacquare i vetrini con 3 cambi di PBS per 2 minuti ciascuno.
5. I vetrini sono ora pronti per la colorazione PCNA.
6. (Facoltativo): Colorazione specifica di tessuti fissati in formalina, inclusi in paraffina può essere evidenziata utilizzando un metodo Recupero di Epitopi mediante calore (Heat Induced Epitope Retrieval) (HIER).
7. (Facoltativo): Se si utilizzano tessuti fissati in alcool, le sezioni di tessuto si possono immergere poi in 10% di NBF per 30-60 secondi. Risciacquare bene. Questo potrebbe migliorare la morfologia.
Per Cellule Coltivate e Sospensioni cellulari
1. Fissare le cellule in 70% di alcool oppure acido-etanolo per 15-20 minuti a 4°C. Si possono utilizzare acetone oppure fissativi Methacarn.
2. Se necessario, bloccare per la perossidasi endogena con 3% H2O2 in metanolo per 10 minuti.
3. Lavare in 3 cambi di PBS per 2 minuti ciascuno.
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Erfahrung
Übersetzungserfahrung in Jahren: 17. Angemeldet bei ProZ.com seit: Aug 2000.
Freelance translator since 1998 , in the following fields: - news
- arts and humanities
- wine and food labels- technical manuals
- textile machinery
- fashion
- furniture
- - pedagogy and psychology, tourism
I am skilled in translations from English/German to Italian. I also translate from Italian into English and have texts revised by native translators. I guarantee for quality translations.
1991 Comparative Pedagogy Erasmus Course in Groningen, The Netherlands
1996 Degree in Foreign Languages and Literature, University of Udine, University of Udine
1999 post-graduate course on Translation techniques
2000 Sommelier course first level
2002 Certificate of Proficency in English Grade "A"
I also teach English and German in secondary school and I hold language courses for adults.
I guarantee for accuracy and speed.
References and detailed CV available on request.
Software: Windows 98 installed MS-Office 97 (Word, Excel, Publisher, Frontpage), Netscape Communicator.
Hardware: PC Pentium II 256 Mb RAM- 12 Gb HD , scanner 1200 dpi , colour printer 720 dpi, modem-fax 56K, Internet ADSL connection
[email protected] phone + 39.0432.566098 (from 9 a.m. to 7 p.m.) if absent please leave a message on answering machine.
mobile phone 39.3478441525
contact me for free quotations
PAYMENT TERMS: 30 DAYS DATE OF INVOICE - BANK
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