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|Human Cell lysate that Human GAP43 / Neuromodulin transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).|
|A DNA sequence encoding the human GAP43 (P17677) (Met 1-Ala 238) was fused with a polyhistidine tag at the C-terminus.|
|The recombinant human GAP43 consists of 249 amino acids and has a predicted molecular mass of 26.2 kDa. As a result of glycosylation, the apparent molecular mass of rhGAP43 is approximately 47 kDa in SDS-PAGE under reducing conditions.|
|Cell lysate was prepared by homogenization 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.|
|Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.|
|12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.|
|Samples are stable for up to twelve months from date of receipt.|
|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. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.|
|1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).|
|Store at 4℃. After re-dissolution, aliquot and store at -80℃.|
|Western blot (WB): Use at an assay dependent dilution.|
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
Neuromodulin, also known as Axonal membrane protein GAP-43, Growth-associated protein 43, Neural phosphoprotein B-50, pp46 and GAP43, is a cell membrane protein which belongs to the neuromodulin family. Neuromodulin / GAP43 contains one IQ domain. Neuromodulin / GAP43 is associated with nerve growth. It is a major component of the motile "growth cones" that form the tips of elongating axons. Neuromodulin / GAP43 is involved in neurite outgrowth, a crucial process for the differentiation of neurons. The sudden infant death syndrome (SIDS) is the main cause of postneonatal infant death and its cause is still unknown. Neuromodulin / GAP43 is a marker of synaptic plasticity and is critical for normal development of the serotonergic innervation. Neuromodulin / GAP43 is a major cortical cytoskeleton-associated and calmodulin binding protein that is widely and abundantly expressed during development, maintained in selected brain structures in the adult, and reinduced during nerve regeneration. CAP23 and GAP43 are functionally related intrinsic determinants of anatomical plasticity. These proteins function by locally promoting subplasmalemmal actin cytoskeleton accumulation.