Technical Data
D3878-16
DNA, ss
Description:
Apoptosis is a genetically and biochemically regulated mechanism of programmed cell death that plays an important role in a variety of human diseases including cancer, immune disorders, and neurologic, cardiovascular and infectious diseases. Decreased stability of apoptotic DNA toward thermal denaturation is induced by the proteolysis of DNA-bound proteins during the execution of apoptosis. This conclusion is supported by the binding of anti-ssDNA monoclonal antibodies to non-apoptotic cells treated with proteinase K before heating and by prevention of DNA denaturation and MAb reactivity in apoptotic nuclei reconstituted with histones (Frankfurt et al. 1996, 1997). Thus, staining of cell suspensions and tissue sections with such antibodies following a heat treatment, which induces DNA denaturation in situ only in apoptotic nuclei, is a specific and sensitive method for the detection of apoptotic cells. D3878-16 provides a cellular marker specific for apoptotic death that is independent of internecleosomal DNA fragmentation and is useful for the detection of different stages of apoptosis in various cell types.

Applications:
Suitable for use in ELISA, Flow Cytometry and Immunohistochemistry and Immunofluorescence. Other applications have not been tested.

Recommended Dilutions:
ELISA: 1:2000-1:5000
Immunohistochemistry (Paraffin): 1:10. May require additional antigen retrieval methods. See attached protocol.
Immunohistochemistry (Frozen): 1:10
Immunofluorescence: 1:10
Flow Cytometry: 1:10-1:200. See attached protocol
Optimal dilutions to be determined by the researcher.

Control: Apoptotic cells

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
MabIgM2Q2282Purified
SizeStorageShippingSourceHost
50ug-20CBlue IceBovineMouse
Concentration:
~0.1mg/ml
Immunogen:
Calf thymus single-stranded DNA.
Purity:
Purified
Form
Supplied as a liquid in PBS, pH 7.4, 5% fetal bovine serum (FBS). Preservative free
Specificity:
Recognizes single-stranded DNA from all species. Specifically recognizes deoxycytidine and requires a stretch of ssDNA of at least 25-30 bases in length for the binding. Does not recognize DNA in double-stranded conformations. Does not recognize necrotic cells.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Costourous J.G., et al. (2004). J Orthop Res. 22:678-683. 2. Tuder, R.M., et al. (2003). Am J Respir Cell Mol. Biol. 29:88-97. 3. Sivagurunathan S., et al. (2002). Can. Res. 62:3868-3875. 4. Gum J.R., et al. (2001). Tumor Biol. 61:3472- 3479. 5. Studzinski D.M. and Benjamins JA. J. (2001). Neuroscience Research 66:691-697.