Product: Anti-BrdU, clone IIB5
Cat. No.: MC-328 (1 mL)
Background:
Bromo-deoxy-Uridine (BrdU) is a thymidine
analogue and when offered to proliferating cells
it is incorporated into reduplicating DNA. The
antibody is specific for DNA in which BrdU has
been incorporated. Immunogen: BrdU coupled
to BSA. In immunoassays this antibody reacts
strongly with free or carrier-protein coupled BrdU
but not with other nucleosides. In
immuncytochemistry the antibody only
recognizes BrdU in denatured (single stranded)
DNA. The BrdU antibody is 100% cross-reactive
with Iodo-Deoxy-Uridine (IrdU). Therefore, IdU
instead of BrdU can be used for in vivo studies.
Specificity:
Reacts with human BrdU.
Species Reactivity:
Human. Others not tested.
Ig Isotype:
Mouse IgG1
Format:
1 mL of diluted ascites, sufficient for 100 tests.
Storage:
Short term: 4°C. For long term, -20°C or below.
Aliquot to avoid freeze/thaw cycles.
Applications and Suggested Dilutions:

Immunohistochemistry: Use on frozen
sections and paraffin embedded tissue after
protease digestion. Dilute in 0.15 M
phosphate buffered saline with 1% BSA and
1% sodium azide. Suggested working
dilution is 1:5–1:10. Optimal dilution should
be tested by serial dilution.

Functional application:
Pulse labeling of dividing cells will allow the
immunocytochemical identification of S-phase
cells. BrdU incorporation can be analyzed in
in vitro cell cultures by adding BrdU to the
tissue culture medium (10μM final conc.).
Exposure periods as brief as 10 minutes allow
sufficient BrdU incorporation for reliable
analysis. For in vivo applications, parenteral
administrations of BrdUrd in dosages of
5mg/kg appear to be effective. BrdU
disappears from the circulation in 30 minutes.
Ex vivo labeling can also be achieved by brief
culturing of small viable tissue specimens,
immediately after removal, in medium
containing 10μM BrdU. In this context the
anti-BrdU antibody IIB5 can be used for:
1. Radioimmunochemical determination of
circulating BrdU levels after parenteral
administration.
2. Detection of S-phase cells in cell
suspensions by immunofluorescence
microscopy.
3. Detection of S-phase cells in tissue
sections by immunoperoxidase or
immunofluorescence methods alone or in
double immunocytochemical staining
approaches.
4. Determination of the percentage of
proliferating cells by flow cytometry
analysis.
5. Quantitative evaluation of the number of
cells in the various phases of the cell
cycle (G
1
,S,G
2
,-M) by dual parameter
flow-cytometry analysis.
The optimal dilution for a specific application
should be determined by the researcher.
Limitations:
For in vitro research use only. Not for use in
diagnostics or in humans.
Warranty:
No warranties, expressed or implied, are made
regarding the use of this product. KAMIYA
BIOMEDICAL COMPANY is not liable for any
damage, personal injury, or economic loss
caused by this product