产品名称

TUNEL Assay试剂盒- BrdU-Red
参阅全部 DNA fragmentation 试剂盒

检测方法

Fluorescent

样品类型

Tissue, Adherent cells, Suspension cells

检测类型

Quantitative

检测时间

3h 00m

种属反应性

与反应:?Mammals, Other species

产品概述

TUNEL Assay Kit – BrdU-Red ab66110 uses a convenient and sensitive method to detect DNA fragmentation by flow cytometry and fluorescence microscopy in live cells.

The TUNEL assay is used to detect DNA fragmentation, such as in apoptosis. It uses?terminal deoxynucleotidyl transferase (TdT) to catalyze the incorporation of?deoxynucleotides at the?free 3′-hydroxyl ends of fragmented DNA. The?deoxynucleotides are then?labeled in a variety of ways for detection of the degree of DNA fragmentation.

This TUNEL assay protocol is based on Br-dUTP (bromolated deoxyuridine triphosphate nucleotide), which can be more readily incorporated into DNA strand breaks by the TdT enzyme than other dUTP labels such as FITC, biotin or dioxigenin. The greater incorporation rate produces a brighter signal when the Br-dUTP sites are detected with an anti-BrdU monoclonal antibody directly labeled with a red fluorochrome.

The BrdU-Red signal can be analyzed at Ex/Em 488/576 nm, with an optional 7-AAD counterstain at Ex/Em 488/655nm.

This TUNEL assay kit includes both negative and positive control cells. It is designed to be suitable for studying DNA fragmentation in GFP-transfected cells.

Tunel assay protocol summary:
– fix cells / tissues with formaldehyde, or deparaffinize and rehydrate if paraffin sections, and wash
– incubate cells in 70% ethanol for 30 min at 4oC, or if tissues incubate with proteinase K solution for 5 min at room temp and refix with formaldehyde
– wash
– incubate in DNA labeling solution for 60 min at 37oC
– wash
– incubate in antibody solution for 30 min at room temp
– add 7-AAD / RNase A solution and incubate for 30 min at room temp
– analyze with flow cytometry or fluorescent microscopy

说明

This product is manufactured by BioVision, an Abcam company and was previously called K404 ApoBrdU Red DNA Fragmentation Kit. K404-60 is the same size as the 60 test size of ab66110.

This kit is BrdU-Red labeled (Ex/Em = 488/576 nm). It was previously called?TUNEL Assay Kit – In situ BrdU-Red DNA Fragmentation.

To use FITC (Ex/Em = 495/519 nm) as a label, we recommend?TUNEL Assay Kit – FITC (ab66108).

For chromogenic TUNEL staining, we recommend?TUNEL Assay Kit – HRP-DAB ab206386.

Find out more about the TUNEL method in the?TUNEL staining / TUNEL assay guide.

Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

平台

Flow cytometer, Fluorescence microscope

性能

存放说明

Please refer to protocols.

组件

标识符

60 tests

7-AAD/RNase Staining Buffer

Amber bottle

1 x 30ml

Anti-BrdU-Red Antibody

Orange

1 x 300μl

Br-dUTP

Violet

1 x 480μl

Negative Control Cells

Neutral

1 x 5ml

Positive Control Cells

Brown

1 x 5ml

Reaction Buffer

Green

1 x 600μl

Rinse Buffer

Red

1 x 120ml

TdT Enzymes

1 x 45μl

Wash Buffer

Blue

1 x 120ml