Human Munc18-1/STXBP1 Baculovirus-Insect cells Overexpression Lysate: Product Information
This Human Munc18-1/STXBP1 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of Munc18-1/STXBP1 protein (Cat: 11751-H20B) from the overexpression lysate was verified.
A DNA sequence encoding the human STXBP1 isoform 1 (P61764-1) (Met 1-Ser 594) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
The recombinant human STXBP1/GST chimera consists of 831 amino acids and has a calculated molecular mass of 95.4 kDa. It migrates as an approximately 80 kDa band in SDS-PAGE under reducing conditions.
Human Munc18-1/STXBP1 Baculovirus-Insect cells Overexpression Lysate: Usage Guide
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human Munc18-1/STXBP1 Baculovirus-Insect cells Overexpression Lysate: Alternative Names
Human MUNC18-1 Overexpression Lysate; Human NSEC1 Overexpression Lysate; Human P67 Overexpression Lysate; Human RBSEC1 Overexpression Lysate; Human UNC18 Overexpression Lysate
Munc18-1/STXBP1 Background Information
Syntaxin-binding protein 1, also known as N-Sec1, Protein unc-18 homolog 1, MUNC18-1 and STXBP1, is a peripheral membrane protein which belongs to theSTXBP / unc-18 / SEC1 family. STXBP1 is an evolutionally conserved neuronal Sec1/Munc-18 (SM) protein that is essential in synaptic vesicle release in several species. It may participate in the regulation of synaptic vesicle docking and fusion, possibly through interaction with GTP-binding proteins. STXBP1 is essential for neurotransmission and binds syntaxin, a component of the synaptic vesicle fusion machinery probably in a 1:1 ratio. It can interact with syntaxins 1, 2, and 3 but not syntaxin 4. STXBP1 may also play a role in determining the specificity of intracellular fusion reactions. Defects in STXBP1 are the cause of epileptic encephalopathy early infantile type 4 (EIEE4). Affected individuals have neonatal or infantile onset of seizures, suppression-burst pattern on EEG, profound mental retardation, and MRI evidence of hypomyelination.
syntaxin binding protein 1
Gengyo-Ando K., et al., 1996, J. Neurosci. 16: 6695-6702.
Swanson D.A., et al., 1998, Genomics 48: 373-376.
The MGC Project Team. 2004, Genome Res. 14: 2121-2127.
Rush J., et al., 2005, Nat. Biotechnol. 23: 94-101.
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