罗氏凋亡试剂盒说明书

For life science research only. Not for use in diagnostic procedures.

In Situ Cell Death Detection Kit, POD

y Version 13.0

Content version: November 2010

Kit for immunohistochemical detection and quantification of apoptosis (pro-grammed cell death) at single cell level, based on labeling of DNA strand breaks (TUNEL technology): Analysis by light microscopy.

Cat. No. 11 684 817 910

Store the kit at 15 to 25°C

1 Kit (50 tests)

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1. reface1.1

Table of contents

1. reface .............................................................................................................................21.1Table of contents .....................................................................................................................................21.2 Kit contents ................................................................................................................................................32. Introduction .....................................................................................................................52.1Product overview .....................................................................................................................................52.2 Background information .......................................................................................................................83. 3.13.2

Procedures and required materials ............................................................................9Flow chart .................................................................................................................................................10Preparation of sample material ........................................................................................................103.2.1Adherent cells, cell smears and cytospin preparations ..............................................113.2.2 Tissue sections ...........................................................................................................................11

3.2.2.1 Treatment of paraffin-embedded tissue ............................................................113.2.2.2Treatment of cryopreserved tissue ......................................................................12

3.3Labeling protocol ...................................................................................................................................13

3.3.1 Before you begin .......................................................................................................................133.3.2Labeling protocol for adherent cells, cell smears, cytospin

preparations and tissues ........................................................................................................143.3.3 Labeling protocol for difficult tissue ..................................................................................15

3.4 Signal conversion ..................................................................................................................................16

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4. Appendix .......................................................................................................................174.1 Troubleshooting .....................................................................................................................................174.2 References ...............................................................................................................................................20P

4.3 Ordering guide .......................................................................................................................................21

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11 684 817 910

http://www.77cn.com.cn

In Situ Cell Death Detection Kit, PODy Version 13.0

1.2 Kit contentsCaution

The Label solution contains cacodylate, toxic by inhalation and swallowed, and cobalt

dichloride, which may cause cancer by inhalation. Avoid exposure and obtain special instructions before use.

When using do not eat, drink or smoke. After contact with skin, wash immediately with plenty of water. In case of accident or if you feel unwell seek medical advice immediately (show label where possible).

Collect the supernatants from the labeling reactions in a tightly closed, non-break-able container and indicate contents. Discard as regulated for toxic waste.

: In contrast to preceding kits/vials, now the Enzyme Solution does no longer contain potassium cacodylate. Thus vial 1 is not toxic.Please refer to the following table for the contents of the kit.Vial/Cap1blue

Label

Contents

Kit contents

Enzyme Solution Terminal deoxynucleotidyl transferase from calf thy-mus (EC 2.7.7.31), recombinant in E. coli, in storage buffer 10× conc. × 50 l 5Label Solution

Nucleotide mixture in reaction buffer 1× conc. 5 × 550 l

Anti-fluorescein antibody, Fab fragment from sheep, conjugated with horse-radish peroxidase (POD) Ready-to-use 3.5ml

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Converter-POD

Additional In addition to the reagents listed above, you have to prepare several solutions. In the equipment requiredtable you will find an overview about the equipment which is needed for the different

procedures.

Detailed information is given in front of each procedure.

Procedure

Adherent cells, cell smears and cytospin preparations (section 3.2.1)

Cryopreserved tissue (section 3.2.2.2)

Equipment

Reagents

Washing buffer: Phosphate buffered saline (PBS*)

Blocking solution: 3% H2O2 in methanol Fixation solution: 4% Paraformalde-hyde in PBS, pH 7.4, freshly prepared Permeabilisation solution: 0.1% Triton X-100 in 0.1% sodium citrate, freshly prepared (6)

Preparation of sample material (section 3.2)

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