SAP102 Overexpression Lysate

Western Blot: SAP102 Overexpression Lysate (Adult Normal) [NBL1-09910] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for SAP102.

• Reactivity: Hu
• Applications: WB

SAP102 Overexpression Lysate Summary

• Description:

  SAP102 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC210860
  
  Accession#: NM_021120
  
  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: DLG3

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: 90.1 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 SAP102 Overexpression Lysate

• discs, large homolog 3 (Drosophila)
• KIAA1232discs, large homolog 3 (neuroendocrine-dlg, Drosophila)
• MRX90MRX
• NEDLGdisks large homolog 3
• neuroendocrine-DLG
• SAP-102
• SAP102NE-Dlg
• Synapse-associated protein 102
• XLMR

Background

Synapse-Associated Protein 102 (SAP102) is one of a family of plasma membrane-associated proteins found in synaptic junctions. Like other members of the family, SAP102 has three ~90 amino acid repeats called PDZ domains followed by an SH3 domain and a yeast guanylate kinase homology (GuK) domain. It is hypothesized that PDZ-domain interactions play a role in receptor and channel clustering which contributes to neuronal plasticity. SAP102 is believed to participate in the clustering of certain proteins, including NMDA receptors, shaker-type potassium channels at the synaptic membrane in CNS neurons. NMDA receptors and Shaker-type potassium channels both share C-terminal sequence homology consisting of a threonine/serine-X-valine-COOH (T/SXV) motif. Other neuronal proteins that share this motif (beta 1 adrenergic receptor, some serotonin receptors, some sodium channel subunits, and additional potassium channel subunits) may interact with SAP102 by binding to its PDZ domains. Neuronal nitric oxide synthase (nNOS), which lacks the T/SXV motif but which has its own PDZ domain, has been shown to associate with SAP102 in vitro through a pseudo-homotypic PDZ-PDZ interaction.

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: DLG3