BrdU antibody

Catalog No. ABIN125982

Product Details anti-BrdU Antibody

Target: BrdU (Bromodeoxyuridine (BrdU))

Reactivity: Please inquire

Host: Mouse

Clonality: Monoclonal

Conjugate: This BrdU antibody is un-conjugated

Application: Flow Cytometry (FACS), Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))

Immunogen: BrdU conjugated to BSA.

Clone: IIB5

Isotype: IgG1

Application Details

Application Notes: Flow Cytometry. In situ-hybridization. Immunohistochemistry on Frozen Sections: Immunohistochemistry on Paraffin Sections: 1/10 for 1h at RT. Proteolytic treatment withpepsin (see Protocol).
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: The Use of Bromodeoxyuridine and Anti-Bbromodeoxyuridine in the Detection of CellProliferation Activity (Labeling and Detection Methods)1) IntroductionThe proliferative activity of cells is estimated by measuring the labeling-index (L. I. ). The L. I. is defined as the fraction of cells in S-phase at the moment of labeling. In the past the L. I. was measured by incorporation of radioactive labeled thymidine into the DNA. With theintroduction of immunohistochemically detectable nucleotides a new non-radioactivemethod for the estimation of the L. I. has become available. The method is based upon theincorporation of bromodeoxyuridine (BrdU), a thymidine analogue, into reduplicating DNA. With an antibody directed against bromodeoxyuridine (BrdU) the labelled cells can beimmunocytochemically detected. Especially on tissue sections this method has manyadvantages. Measurement of the proliferative activity of e. g. tumour cells is now within thereach of each laboratory. 2) Labeling with bromodeoxyuridinea) In vitro labelling: Cell cultures. Addition of BrdU (10 µM final concentration) to the cell culture medium. After 10 to 30minutes the cells are harvested and fixed in the appropriate fixative. b) In vivo labelling. Animals are injected intraperitoneally with 5-50 mg BrdU/kg bodyweight. After 1h theanimals are sacrificed, the organsremoved, and fixed or snap frozen. c) Ex vivo labeling. )i) 'Single' cell biopsies. - Aspiration biopsies e. g. : Bone marrow- Brush preparations e. g. : Lung- Biopsies are transferred immediately to a cell culture tube (15 ml) containing 10 mlmedium pre-incubated at 37°C. Medium: RPMI 1640 (Hepes buffered)10% Foetal calf serum10 µM BrdU¦Incubation one hour at 37°C. After labeling transfer biopsy to the appropriate fixative. ii) Solid tissue biopsies. Small biopsies, 1—3 mm3, are taken and immediately transferred to a cell culture tube (15ml) containing 10 ml medium pre-incubated at 37°C. (see 2. C1). Add three drops of 30%H2O2, and firmly close screw cap. Incubate one hourat 37°C. After incubation, transfer biopsy to the appropriate fixative. 3) Fixation of biopsiesOptions: - Frozen sections- Cytospin preparations- Flow cytometry- Paraffin sectionsFrozen Sections: After labeling with BrdU, specimens are snap frozen in isopentanequenched in liquid nitrogen. Sections (2-5 µm) are cut in a cryostat at -20°C, and mounted

Restrictions: For Research Use only

Handling

Format: Liquid

Handling Advice: Avoid repeated freezing and thawing.

Storage: 4 °C/-20 °C

Storage Comment: Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.

Target Details for BrdU

Target: BrdU (Bromodeoxyuridine (BrdU))

Abstract: BrdU Products

Target Type: Chemical

Background: Bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) is a synthetic nucleoside which is an analogue of thymidine. BrdU is commonly used in the detection of proliferating cells in living tissues. BrdU can be incorporated into the newly synthesized DNA of replicating cells (during the S phase of the cell cycle), substituting for thymidine during DNA replication. Antibodies specific to BrdU can then be used to detect the incorporated chemical, thus indicating cells that were actively replicating their DNA. Binding of the antibody requires denaturation of the DNA by heat or acid.