Product nameHuman RBM3 knockout HEK-293T cell line
DescriptionRBM3 KO HEK-293T cell line
Parental Cell LineHEK293T
Mutation descriptionKnockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 2
Knockout validationSanger Sequencing
Recommended control: Human wild-type HEK293T cell line (ab255449). 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% FBS
Initial handling guidelines: Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for 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-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x 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-warmed culture medium and count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- 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.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Number of cells1 x 106 cells/vial, 1 mL
STR AnalysisAmelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
Storage instructionsShipped on Dry Ice. Store in liquid nitrogen.
Storage bufferConstituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether
FunctionCold-inducible mRNA binding protein that enhances global protein synthesis at both physiological and mild hypothermic temperatures. Reduces the relative abundance of microRNAs, when overexpressed. Enhances phosphoryaltion of translation initiation factors and active polysome formation.
Sequence similaritiesContains 1 RRM (RNA recognition motif) domain.
modificationsArg-105 is dimethylated, probably to asymmetric dimethylarginine.
Cellular localizationNucleus. Cytoplasm. Cell projection > dendrite. Localizes in mRNA granules in dentrites.
- Information by UniProt
KO cell lysates
All lanes : Anti-RBM3 antibody [EPR6061(2)] (ab134946) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : RBM3 Knockout HEK-293T cell lysate
Lane 3 : Hela cell lysate
Lane 4 : LnCap cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 18 kDa why is the actual band size different from the predicted?
ab134946 was shown to react with RBM3 in wild-type HEK-293T cells in Western blot with loss of signal observed in RBM3 knockout cell line ab266663 (RBM3 knockout cell lysate ab258170). Wild-type HEK-293T and RBM3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab134946 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
Homozygous: 2 bp deletion in exon 2
ab266663 has not yet been referenced specifically in any publications.