TdT

This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.

Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

Application Name	Verified	Min Dilution	Max Dilution
Immunofluorescence

Instructions For Use

OBT0009

Standard bone marrow or peripheral blood smears may be used for immunofluorescence staining. Cytocentrifuge preparations of cell suspensions made up in RPMI-1640 tissue culture medium containing 10% foetal calf serum are also suitable. Frozen sections should be cut as thin as possible. Store unfixed slide preparations in a dessicator at room temperature.

1. Circle two areas of interest (1 cm diameter) with diamond pencil on the slide to be stained. One will be used as a control area and the other is for the test. Fix smears for 30 min. in absolute methanol at 4^oC immediately before staining. Air dry.

2. Block by adding a drop of normal goat serum to both circled areas. Let stand for 5 min. Rinse off goat serum with PBS.

3. Apply 25 ul rabbit anti human TdT antibody mixture to the test area and 25 ul of negative control to control area and allow to react for 30 min in a humid chamber.

4. Rinse avoiding cross-contamination of reagents. Now wash for 10 min. in a Coplin jar with several changes of PBS. Agitate occasionally for thorough washing.

5. Wipe the slide, avoiding cells, and shake excess wash solution off circled area. Spread 25 ul of FITC goat anti rabbit IgG F(ab')₂ and incubate in humid chamber for 30 min in the dark.

6. Rinse off and wash for 10 min. in a Coplin jar with several changes of PBS.

7. Apply small drops of glycerol/PBS (mounting medium) to two glass cover slips and invert circled areas on slide over dropped cover slips.

8. Examine by epifluorescence and phase microscopy. Evaluate the percentage of nucleated cells exhibiting nuclear fluorescence.

Synonyms

Terminal Deoxynucleotidyl Transferase

UniProt

P04053

Entrez Gene

DNTT

GO Terms

GO:0006260 DNA replication

GO:0003677 DNA binding

GO:0003887 DNA-directed DNA polymerase activity

GO:0003912 DNA nucleotidylexotransferase activity

GO:0046872 metal ion binding

GO:0005634 nucleus

GO:0006304 DNA modification

OBT0009

156783 161330 164635 166152 168814

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