OverviewProduct nameHuman LMNB1 (Lamin B1) knockout HeLa cell lineParental Cell LineHeLaOrganismHumanMutation descriptionKnockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 1 and 2 bp insertion in exon 1Passage numberKnockout validationSanger Sequencing, Western Blot (WB)Tested applicationsSuitable for:WBmore detailsBiosafety level2General notesRecommended control: Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.Cryopreservation cell medium:Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.Culture medium:DMEM (High Glucose) + 10% FBSInitial handling guidelines:Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80ºC. Storage at -80ºC may result in loss of viability.1. Thaw the vial in 37ºC water bath approximately 1-2 minutes.2. Transfer the cell suspension (0.8 ml) to a 15 ml/50 ml conical sterile polypropylene centrifuge tube containing 8.4 ml pre-warmedculture medium, wash vial with an additional 0.8 mlculture medium(total volume 10 ml) to collect remaining cells, and centrifuge at 201x g(rcf) for 5 minutes at room temperature. 10 ml represents minimum recommended dilution. 20 ml represents maximum recommended dilution.3. Resuspend the cell pellet in 5 ml pre-warmedculture mediumand count using a haemocytometer (Click here to view haemocytometer protocol) or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x104cells/cm2. This should allow for confluency within 48 hours. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.4. Incubate the culture at 37ºC incubator with 5% CO2. Cultures should be monitored daily.Subculture guidelines:All seeding densities should be based on cell counts gained by established methods.A guide seeding density of 2x104cells/cm2is recommended for confluency (80-90% confluence) within 48 hours.A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.Cells should be passaged when they have achieved 80-90% confluence.Click here to view the Mammalian cell tissue culture protocolThis product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.PropertiesNumber of cells1 x 106 cells/vial, 1 mLViability~90%Adherent /SuspensionAdherentTissueCervixCell typeepithelialDiseaseAdenocarcinomaGenderFemaleSTR AnalysisAmelogenin XD5S818: 11, 12D13S317: 12, 13.3D7S820: 8, 12D16S539: 9, 10vWA: 16, 18TH01: 7TPOX: 8, 12CSF1PO: 9, 10Mycoplasma freeYesStorage instructionsShipped on Dry Ice. Store in liquid nitrogen.Storage bufferConstituents: 8.7% DMSO, 2% Cellulose, methyl etherResearch areasCell BiologyApoptosisNucleusLaminsTags & Cell MarkersSubcellular MarkersNucleusNuclear EnvelopeSignal TransductionCytoskeleton / ECMCytoskeletonIntermediate FilamentsClass VLaminsCancerCell DeathApoptosisNucleusLaminsTargetFunctionLamins are components of the nuclear lamina, a fibrous layer on the nucleoplasmic side of the inner nuclear membrane, which is thought to provide a framework for the nuclear envelope and may also interact with chromatin.Involvement in diseaseDefects in LMNB1 are the cause of leukodystrophy demyelinating autosomal dominant adult-onset (ADLD) [MIM:169500]. ADLD is a slowly progressive and fatal demyelinating leukodystrophy, presenting in the fourth or fifth decade of life. Clinically characterized by early autonomic abnormalities, pyramidal and cerebellar dysfunction, and symmetric demyelination of the CNS. It differs from multiple sclerosis and other demyelinating disorders in that neuropathology shows preservation of oligodendroglia in the presence of subtotal demyelination and lack of astrogliosis.Sequence similaritiesBelongs to the intermediate filament family.Post-translationalmodificationsB-type lamins undergo a series of modifications, such as farnesylation and phosphorylation. Increased phosphorylation of the lamins occurs before envelope disintegration and probably plays a role in regulating lamin associations.Cellular localizationNucleus inner membrane.Target information above from: UniProt accessionP20700The UniProt ConsortiumThe Universal Protein Resource (UniProt) in 2010Nucleic Acids Res. 38:D142-D148 (2010).Information by UniProt