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Frontier™ Chemiluminescent Bromodeoxyuridine (BrdU) Cell Proliferation ELISA Kit
Exalpha's Frontier™ BrdU Proliferation Assay is a completely HTS compatible, non-isotopic, chemiluminescent 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
- Chemiluminescent assay
- HTS compatible format
- Optional spin step
- High sensitivity
- Best Signal to Noise ratio available
- 4 deg C storage with long shelf life
- Non-radioactive
- 2 hour protocol
- Suitable for all species
- Suitable adherent or non-adherent cell types

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

Ordering Information

Cat. No.
X1623K1
X1623K2
X1623K3

Size
200 Tests
1000 Tests
5000 Tests

For larger quantity pricing, please contact Customer Service

 

Protocol Summary: Not to be used in place of detailed protocol. See kit manual for complete 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 chemiluminescent substrate
15. Read • Read immediately (reaction may be read for up to 30 minutes).