Protein Name & Synonyms:
Immunohistology - Frozen, Flow Cytometry, Immunohistology - Paraffin
IgG concentration 1.0mg/ml
- FLOW CYTOMETRY ANALYSIS
Prepare the following solutions before proceeding:
Phosphate buffered saline (PBS)
2N HCl containing 0.5% Triton X-100
PBS containing 0.05% Tween-20
PBS containing 1% BSA (PBS/BSA)
10mg/ml Propidium iodide (PI)
0.1M Na2B4O7, pH 8.5
1. Add BrdU to the cell suspension in culture medium to a final concentration of 10 Î¼mol/L and incubate for 30 minutes in a CO2 incubator at 37°C.
2. Wash cells twice with PBS/BSA by centrifuging at 500g for 10 minutes, decant supernatant and resuspend in a minimum volume of PBS.
3. Add cells slowly into 5ml of 70% ethanol at -20°C, mixing continuously (vortex preferred).
Incubate on ice for 30 minutes.
4. Centrifuge at 500g for 10 minutes, decant supernatant, and resuspend cell pellet.
5. Add 2ml of 2N HCl containing 0.5% Triton X-100 and incubate the cells for 30 minutes at room temperature (preferably on a rocking platform).
6. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend in 3 ml of 0.1M Na2B4O7, pH 8.5 .
7. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend the cells in PBS/BSA + 0.05% Tween-20. Adjust cell concentration to 1 x 107/ml.
8. Aliquot 100ul of cell suspension into required number of 12 x 75mm tubes.
9. Incubate the cells with the BrdU antibody at the recommended dilution for 45 minutes at room temperature or overnight at 4°C.
10. Add 2 ml of PBS/BSA and centrifuge the cells at 1000rpm for 5 minutes.
11. If a secondary antibody layer is required then decant the supernatant and
This product recognizes the thymidine analogue bromodoxyuridine (BrdU), which can be incorporated into DNA during S-phase of the cell cycle. The BU20a antibody is suitable for detecting incorporated BrdU in a wide variety of cell types and is suitable for use on tissue sections in double-labelling techniques.
Store at +4 °C or at -20 °C 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.
If stored in this manner, this product is stable for 18 months after receipt.
Purified IgG - liquid
Phosphate buffered saline
Bromodeoxyuridine conjugated to BSA.
Spleen cells from immunised Balb/c mice were fused with cells of the NS1 myeloma cell line.