F-Spondin/SPON1 Overexpression Lysate

Western Blot: Spondin-1 Overexpression Lysate (Adult Normal) [NBL1-16417] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Spondin-1.

• Reactivity: Hu
• Applications: WB

F-Spondin/SPON1 Overexpression Lysate Summary

• Description:

  F-Spondin/SPON1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC218131
  
  Accession#: NM_006108
  
  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: SPON1

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: 90.8 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 F-Spondin/SPON1 Overexpression Lysate

• FSpondin
• F-Spondin
• KIAA0762VSGP/F-spondin
• MGC10724
• SPON1
• spondin 1, (f-spondin) extracellular matrix protein
• spondin 1, extracellular matrix protein
• spondin-1
• Vascular smooth muscle cell growth-promoting factor
• VSGP

Background

SPON1 is a member of a subgroup of the thrombospondin type 1 (TSR) class molecules, defined by two domains of homology, the FS1/FS2 and TSR domains. The TSRs of SPON1 proteins are typical of class 2 TSRs. SPON1, which is similar to thrombospondin, is a extracellular matrix attached molecule that promotes neurite outgrowth and inhibits angiogenesis. Analysis of gain and loss of function experiments reveal that SPON1 is required for accurate pathfinding of embryonic axons, and plays a dual role in patterning axonal trajectories. It promotes the outgrowth of commissural and inhibits the outgrowth of motor axons, and has also been implicated in inflammatory processes in the nervous system.

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: SPON1