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Rabbit anti Human TdT Kit

Product Type
Kits
Isotype
Polyclonal IgG
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
OBT0009 IF 20 Tests
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.

Product Details

Target Species
Human
Product Form
Kit
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)

Storage Information

Storage
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.

More Information

UniProt
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
Regulatory
For research purposes only

Applications of TdT

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
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.