|Baculovirus-Insect Cell lysate that Human CASK Kinase 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 CASK (O14936-4) (Ala 2-Tyr 898) was expressed and purified with two additional amino acids (Gly & Pro) at the N-terminus.|
|The secreted recombinant human CASK consists of 899 amino acids and predicts a molecular mass of 102.1 KDa. The apparent molecular mass of the protein is approximately 102 KDa in SDS-PAGE under reducing conditions due to glycosylation.|
|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.
Peripheral plasma membrane protein CASK, also known as calcium/calmodulin-dependent serine protein kinase, CASK and LIN2, is a nucleus, cytoplasm and cell membrane protein which belongs to the MAGUK family. CASK / LIN2 contains one guanylate kinase-like domain, two L27 domains, one PDZ (DHR) domain, one protein kinase domain and one SH3 domain. CASK / LIN2 is ubiquitously expressed. Expression of CASK / LIN2 is significantly greater in brain relative to kidney, lung, and liver and in fetal brain and kidney relative to lung and liver. CASK / LIN2 is a multidomain scaffolding protein with a role in synaptic transmembrane protein anchoring and ion channel trafficking. CASK / LIN2 contributes to neural development and regulation of gene expression via interaction with the transcription factor TRB1. It binds to cell-surface proteins, including amyloid precursor protein, neurexins and syndecans. CASK / LIN2 may mediate a link between the extracellular matrix and the actin cytoskeleton via its interaction with syndecan and with the actin/spectrin-binding protein 4.1. Defects in CASK are the cause of mental retardation X-linked CASK-related (MRXCASK). Mental retardation is characterized by significantly below average general intellectual functioning associated with impairments in adaptative behavior and manifested during the developmental period. Defects in CASK are also the cause of FG syndrome type 4 which is an X-linked disorder characterized by mental retardation, relative macrocephaly, hypotonia and constipation.