BrdU Proliferation
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Bromodeoxyuridine (BrdU) Cell Proliferation ELISA Kit

BrdU Proliferation Assay is a completely HTS compatible, non-isotopic, colorimetric proliferation enzyme-linked immunosorbent assay (ELISA) assay kit for the detection of bromodeoxyuridine incorporation into newly synthesized DNA of adherent and non-adherent cells.

Features:

- Colorimetric assay
- HTS compatible format
- Optional spin step
- High sensitivity
- 4 deg C storage with long shelf life
- Non-radioactive
- 2.5 hour protocol
- Suitable for all species
- Suitable adherent or non-adherent cell types

Performance Summary:

- Sensitivity: 40 cells/well
- Intra-assay C.V.'s: <10% (spin protocol)
- Inter-assay C.V.'s: <10% (spin protocol)

Ordering Information:

Cat. No.
 X1327K1= 200 tests
X1327K2 = 1000 tests
X1327K3 = 5000 tests

For larger quantity pricing, please contact Customer Service


Legend:
BrdU assay. Detection of variable numbers of Jurkat (non-adherent) or CHO cells (adherent) per well. Y axis-left, OD 450-550nm. Y axis-right, signal-to-noise ratio.

Protocol Summary: Not to be used in place of detailed protocol:

1. Cell Plating – no Test Reagent/Drug
(skip step 3 below)		 • Seed cells at 1-2 x 105 cells/ml, 100 ml/well
2. Cell Plating – withTest Reagent/Drug
(see below step 3) 		• Seed cells at 0.5-4 x 105 cells/ml, 50 ml/well
3. Addition of Test Reagent(s)/Drug • Add 50 ml/well, 2X concentration desired
4. Addition of BrdU •  Dilute 500X stock BrdU, add 20 ml/well
(be sure to include a No BrdU control)
5. Incubate • 2-24 hours
6. Fix and Denature
- Adherent Cells
 
• Aspirate (or flick) the media from the cell wells
• Add 200 ml/well Fixing Solution
• Incubate 30 minutes at Room Temp.
• Aspirate the Fixing Solution and blot the plates dry.
- Suspension Cells
No-Spin Procedure		 • Add 200 ml/well Fixing Solution on top of the cells.
• Incubate 1 hour at Room Temp
• Aspirate the Fixing Solution and blot the plates dry.
- Suspension Cells
Spine Procedure		 • Spin the plates for 5 minutes at 1000 rpm.
• Aspirate media, add 200 ml/well Fixing Solution.
• Incubate for 30 minutes, room temp.
• Aspirate the Fixing Solution and blot the plates dry.
7. Wash Step • Wash X3 with 1X wash buffer and blot dry.
8. Detector Antibody • Add 100 ml/well of diluted detector antibody.
9. Incubate • 1 hour at room temp.
10. Wash Step • Wash X3 with 1X wash buffer and blot dry.
11. Conjugate Addition • Add 100 ml/well HRP-conjugate
12. Incubate • Incubate for 30 minutes at room temperature.
13. Wash Step and Final Water Wash • Wash as above. Perform a final distilled water wash by flooding the entire plate with distilled water. Pat dry on absorbent paper towels.
14. Development
 		 • Add 100 ml/well TMB Peroxidase substrate
15. Incubate • 30 minutes at room temperature in the dark.
16. Stop • Add 100 ml of acid Stop Solution to every well
17. Read • Read the at 450/550 nm