Rabbit anti Human TdT kit is a set of reagents intended to allow the identification of Terminal deoxynucleotidyl transferase positive (TdT+) cells by combined fluorescence and phase contrast microscopy.
- Target Species
- Product Form
- Preservative Stabilisers
- All components contain 0.1% Sodium Azide (NaN3)
Components 2, 2C and 3 contain 1% Bovine Serum Albumin
Component M contains 80% glycerol
- Reagents in the Kit
- Kit contains ready-to-use reagents sufficient for twenty slide tests:
1 Normal goat serum - 1.0ml
2 Rabbit anti- human -TdT antibody mixture - 0.5ml
2C Negative control antibody - 0.5ml
3 FITC-goat anti- rabbit IgG F(ab')2 - 1.0ml.
M Mounting medium (80% glycerol in PBS/azide) - 1.0ml
- Additional Reagents Required
- Cold absolute methanol
Phosphate buffered saline (PBS)
- Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
- Shelf Life
- Please see label for expiry date.
- P04053 Related reagents
- Entrez Gene
- DNTT Related reagents
- 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
- For research purposes only
Applications of TdT
|Application Name||Verified||Min Dilution||Max Dilution|
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Instructions For Use
- 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 4oC 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.
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