Preparation And Storage
Recommended Assay Procedures
Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml
Bioimaging:
Methanol Procedure for a 96 well plate:
Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS. Image sample.
Triton-X 100 Procedure for a 96 well plate:
Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS. Image sample.
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
The S100 multigene family consists of a number of small dimeric molecules around 100 residues in length. The original members of this family, S100A and S100B, were initially identified in bovine brain. Other S100 proteins have also been identified, such as S100P from human placenta and S100L from bovine lung. The S100 family consists of at least 10 members which show a cell-type-specific expression pattern. Most S100 proteins are believed to be involved in Ca2+ signaling pathways. S100P consists of 95 amino acids and shares about 50% sequence identity with S100A and S100B, while S100L has been shown to possess 43-47% homology to S100A and S100B. S100B is expressed in the nervous system by glial cells, such as astrocyte, oligodendroglial, ependymal, and microglial cells. The dimeric form of S100B has been implicated in neuronal survival, neurite extension, and astrocyte growth. In addition, S100B expression is induced by stimulations that activate p53, and S100B activity may be involved in PKC-dependent p53 nuclear translocation in early G1 phase. Thus, S100B plays a role in calcium signaling pathways that regulate cell differentiation and cell cycle progression.
Development References (2)
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Jiang H, Shah S, Hilt DC. Organization, sequence, and expression of the murine S100 beta gene. Transcriptional regulation by cell type-specific cis-acting regulatory elements. J Biol Chem. 1993; 268(27):20502-20511. (Biology). View Reference
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Scotto C, Delphin C, Deloulme JC, Baudier J. Concerted regulation of wild-type p53 nuclear accumulation and activation by S100B and calcium-dependent protein kinase C. Mol Cell Biol. 1999; 19(10):7168-7180. (Biology). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.
