Human RBPMS knockout HeLa cell line
- Advanced Validation
- What is this?
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RBPMS KO cell line available to order. KO validated by. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 44 bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
FLJ32971, Heart and RRM expressed sequence, Hermes, RBPMS_HUMAN, RNA binding protein gene with multiple splicing, RNA-binding protein with multiple splicing
- WB
Lab
Western blot - Human RBPMS knockout HeLa cell line (AB264697)
Lanes 1 - 4 : Merged signal (red and green). Green - ab194213 observed at 30 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab194213 was shown to react with RBPMS in wild-type HeLa cells in western blot. Here no band was observed in the RBPMS knockout cell line ab264697 (RBPMS knockout cell lysate ab258631) but other antibodies have shown bands in this lysate below 30 kDa that may represent truncated forms and cleaved fragments. This has not been investigated further. HeLa wild-type and RBPMS knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab194213 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° 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.
Lanes 1 - 4:
Western blot - Anti-RBPMS antibody (<a href='/en-us/products/primary-antibodies/rbpms-antibody-ab194213'>ab194213</a>) at 1/1000 dilution
Lanes 1 - 4:
Western blot - Anti-RBPMS antibody (<a href='/en-us/products/primary-antibodies/rbpms-antibody-ab152101'>ab152101</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
RBPMS Knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human RBPMS knockout HeLa cell line (ab264697)
Lane 3:
RT-4 cell lysate at 20 µg
Lane 4:
U-251 MG cell lysate at 20 µg
Predicted band size: 22 kDa
Observed band size: 30 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RBPMS knockout HeLa cell line (AB264697)
Allele-1 : 1 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human RBPMS knockout HeLa cell line (AB264697)
Allele-2 : 44 bp deletion in exon 2.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
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 seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate 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. 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 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
RBPMS plays a role in the regulation of gene expression by interacting with other proteins to form ribonucleoprotein complexes. These complexes are important for the post-transcriptional management of gene expression impacting mRNA splicing and localization. RBPMS also exhibits involvement in the development of neuronal cells where it contributes to neuronal differentiation and growth by aiding in the precise processing of specific mRNA transcripts that are essential for neural functions.
Pathways
RBPMS participates in the regulation of the mRNA splicing and transport pathways. It interacts with the spliceosome complex influencing alternative splicing events that generate diverse protein isoforms. RBPMS is associated with proteins such as SMN1 which also plays a part in mRNA processing and is integral to the spliceosomal machinery. These interactions make RBPMS a pivotal mediator in translating genetic information into functional proteins therefore affecting cellular growth and differentiation.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Primary Antibodies
AB309094
Anti-RBPMS antibody [EPR27130-21]
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![Flow Cytometry (Intracellular) - Anti-RBPMS antibody [EPR27130-21] (AB309094)](https://content.abcam.com/products/images/rbpms-antibody-epr27130-21-ab309094--flow-cytometry-intracellular-img409585.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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