Preparation and Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
Id proteins were originally characterized as inhibitors of DNA binding and cell differentiation. Id1 through 4 contain an evolutionarily conserved helix-loop-helix (HLH) sequence which is critical for protein-protein interaction(s). Most HLH transcription factors contain a basic amino acid region adjacent to the HLH sequence, the bHLH sequence, which is responsible for DNA binding. bHLH transcription factors fall into 2 major groups designated class A factors, e.g.,E2.2 and E47, and class B factors, e.g., MyoD and myogenin. In vitro studies demonstrate distinct interaction(s) between Id proteins and bHLH transcription factors. While Id proteins contain an HLH domain, they lack the basic region which is required for DNA binding. Therefore, Id proteins are negative regulators of transcription since complexes which contain them do not bind DNA. Id proteins are variably expressed throughout the cell cycle and are regulated by phosphorylation by cyclin-cdk complexes. Thus, Id proteins play an important role in transcriptional regulation of cell cycle related genes. Overexpression of Id3 can induce apoptosis in serum-starved fibroblasts by a mechanism which is correlated to Id3-mediated cell cycle progression and is rescued by over expression of the antiapoptotic proteins Bcl-2 and Bcl-XL. Clone B72-1 recognizes mouse Id3. It does not cross react with mouse Id1 or mouse Id2. A fusion protein containing full length mouse Id3 was used as immunogen. The antibody is routinely tested by western blotting of the Id3-fusion protein. The Id3-fusion protein is observed at 43 kDa (fusion protein MW ~28 kDa and Id3 MW is ~13 kDa).
Development References (7)
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Cooper CL, Brady G, Bilia F, Iscove NN, Quesenberry PJ. Expression of the Id family helix-loop-helix regulators during growth and development in the hematopoietic system. Blood. 1997; 89(9):3155-3165. (Biology). View Reference
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Hara E, Hall M, Peters G. Cdk2-dependent phosphorylation of Id2 modulates activity of E2A-related transcription factors. EMBO J. 1997; 16(2):332-342. (Biology). View Reference
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Kadesch T. Consequences of heteromeric interactions among helix-loop-helix proteins. Cell Growth Differ. 1993; 4(1):49-55. (Clone-specific: Inhibition). View Reference
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Langlands K, Yin X, Anand G, Prochownik EV. Differential interactions of Id proteins with basic-helix-loop-helix transcription factors. J Biol Chem. 1997; 272(32):19785-19793. (Biology). View Reference
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Norton JD, Atherton GT. Coupling of cell growth control and apoptosis functions of Id proteins. Mol Cell Biol. 1998; 18(4):2371-2381. (Clone-specific: Apoptosis). View Reference
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Norton JD, Deed RW, Craggs G, Sablitzky F. Id helix-loop-helix proteins in cell growth and differentiation. Trends Cell Biol. 1998; 8(2):58-65. (Biology). View Reference
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Riechmann V, van Crüchten I, Sablitzky F. The expression pattern of Id4, a novel dominant negative helix-loop-helix protein, is distinct from Id1, Id2 and Id3. Nucleic Acids Res. 1994; 22(5):749-755. (Biology). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
