Description

Nori Human S100B ELISA Kit Summary

Alternative names for S100B: S100 calcium-binding protein B, S100-B,

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

S100 calcium binding protein B (S100B) is a protein of the S-100 protein family. S100B is glial-specific and is expressed primarily by astrocytes, but not all astrocytes express S100B. It has been shown that S100B is only expressed by a subtype of mature astrocytes that ensheath blood vessels and by NG2-expressing cells.^[1] This protein may function in neurite extension, proliferation of melanoma cells, stimulation of Ca²⁺ fluxes, inhibition of PKC mediated phosphorylation, astrocytosis and axonal proliferation, and inhibition of microtubule assembly. In the developing CNS it acts as a neurotrophic factor and neuronal survival protein. In the adult organism it is usually elevated due to nervous system damage, which makes it a potential clinical marker. Altered expression of S100B has been implicated in several neurological, neoplastic, and other types of diseases, including Alzheimer’s disease, Down’s syndrome, epilepsy, amyotrophic lateral sclerosis, melanoma, and type I diabetes. It has been suggested that the regulation of S100B by melittin has potential for the treatment of epilepsy. Serum levels of S100B increase in patients during the acute phase of brain damage. S100B levels have been reported to rise prior to any detectable changes in intracerebral pressure, neuroimaging, and neurological examination findings. Thus, the major advantage of using S100B is that elevations in serum or CSF levels provide a sensitive measure for determining CNS injury before gross changes develop, enabling timely delivery of crucial medical intervention before irreversible damage occurs. S100B serum levels are elevated before seizures suggesting that BBB leakage may be an early event in seizure development. ^[2] An extremely important application of serum S100B testing is in the selection of patients with minor head injury who do not need further neuroradiological evaluation, as studies comparing CT scans and S100B levels have demonstrated S100B values below 0.12 ng/mL are associated with low risk of obvious neuroradiological changes.^[3] In addition, S100B, which is also present in porcine melanocytes, is a reliable marker for melanoma malignancy both in bioptic tissue and in serum.^[4][5]

References

1. Wang DD, Bordey A (2008).   Neurobiol.86 (4): 342–67.
2. Marchi N, et al. (2007). Epilepsia 48 (4): 732–42.
3. Zongo, D., et al. (2012). Annals of Emergency Medicine 59(1): 209–218.
4. Michetti F, et al. (2012). J. Neurochem. 120 (5): 644–59.
5. Cocchia D, et al. (1981). Nature 294 (5836): 85–7.

Product Citations