S100A1 Overexpression Lysate

Western Blot: S100A1 Overexpression Lysate (Adult Normal) [NBL1-15647] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for S100A1.

• Reactivity: Hu
• Applications: WB

S100A1 Overexpression Lysate Summary

• Description:

  S100A1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC200278
  
  Accession#: NM_006271
  
  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: S100A1

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: 10.4 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 S100A1 Overexpression Lysate

• protein S100-A1
• S100 alpha
• S100 calcium binding protein A1
• S100 calcium-binding protein A1S100
• S-100 protein alpha chain
• S-100 protein subunit alpha
• S100 protein, alpha polypeptide
• S100A
• S100A1
• S100-alpha

Background

S100 belongs to the family of calcium binding proteins such as calmodulin and troponin C. S100 proteins are localized in the cytoplasm and/or nucleus of a wide range of cells, and involved in the regulation of a number of cellular processes such as cell cycle progression and differentiation. S100 genes include at least 13 members which are located as a cluster on chromosome 1q21. S100A is composed of an alpha and beta chain whereas S100B is composed of two beta chains. S100 protein may function in stimulation of Ca2+ induced Ca2+ release, inhibition of microtubule assembly, and inhibition of protein kinase C mediated phosphorylation. Reduced expression of this protein has been implicated in cardiomyopathies. Immunoreactive S100 protein localizes in the cytoplasm and nuclei of astrocytes, Schwann's cells, ependymomas and astrogliomas. Ganglion cells do not stain for S100 protein. S100 can be detected in almost all benign naevi and malignant melanocytic tumors of the skin, and in Langerhans cells in the skin.

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: S100A1