S100B Overexpression Lysate

Western Blot: S100B Overexpression Lysate (Adult Normal) [NBL1-15661] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for S100B.

• Reactivity: Hu
• Applications: WB

S100B Overexpression Lysate Summary

• Description:

  S100B Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC200277
  
  Accession#: NM_006272
  
  Protein Tag: C-MYC/DDK
  
  You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: S100B

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 10.5 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for S100B Overexpression Lysate

• beta (neural)
• NEF
• S100 beta
• S100 calcium binding protein B
• S100 calcium-binding protein B
• S100 calcium-binding protein, beta (neural)
• S-100 calcium-binding protein, beta chain, 10protein S100-B
• S-100 protein beta chain
• S-100 protein subunit beta
• S100
• S100B
• S100beta

Background

S100B is a zinc- and calcium-binding protein belonging to the S100 protein family within the EF-hand (helix E-loop-helix F) subgroup (1,2). S100B plays a role in normal central nervous system development, is associated with various neurological diseases such as Alzheimer's and Parkinson's, and it serves as a marker for brain injury (1,2). The S100B protein has a homodimeric structure comprised of two 91-amino acid polypeptide monomers each with a theoretical molecular weight of 10.5 kDa (1,2). Furthermore, each monomer contains two EF-hand regions, four helixes, and a hinge region (2). S100B is predominately expressed in astrocytes, oligodendrocytes, and Schwann cells, but also other cell types including adipocytes (1,3). S100B interacts with toll-like receptor 4 (TLR4) and receptor for advanced glycation end products (RAGE), initiating downstream signaling cascades and transcription factors including JNK/JUN, NFkappaB, and p38, leading to caspase and proinflammatory cytokine production (2). Overall outcomes include neuronal apoptosis, neuroinflammation, and neurodegeneration (1,2). S100B is the most commonly studied astroglia and blood brain barrier biomarker in traumatic brain injury (TBI) (3,4). The serum levels of S100B in patients with TBI is indicative of patient outcomes, where high levels correlate with injury severity and mortality (4). S100B is often in used in combination with additional biomarkers such as glial fibrillary acidic protein (GFAP) and ubiquitin c-terminal hydrolase L1 (UCH-L1) (3,4).

References

1. Yardan, T., Erenler, A. K., Baydin, A., Aydin, K., & Cokluk, C. (2011). Usefulness of S100B protein in neurological disorders. JPMA. The Journal of the Pakistan Medical Association, 61(3), 276-281.

2. Langeh, U., & Singh, S. (2021). Targeting S100B Protein as a Surrogate Biomarker and its Role in Various Neurological Disorders. Current neuropharmacology, 19(2), 265-277. https://doi.org/10.2174/1570159X18666200729100427

3. Thelin, E. P., Nelson, D. W., & Bellander, B. M. (2017). A review of the clinical utility of serum S100B protein levels in the assessment of traumatic brain injury. Acta neurochirurgica, 159(2), 209-225. https://doi.org/10.1007/s00701-016-3046-3

4. Wang, K. K., Yang, Z., Zhu, T., Shi, Y., Rubenstein, R., Tyndall, J. A., & Manley, G. T. (2018). An update on diagnostic and prognostic biomarkers for traumatic brain injury. Expert review of molecular diagnostics, 18(2), 165-180. https://doi.org/10.1080/14737159.2018.1428089

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: S100B