Human RANBP2 knockout HeLa cell line
- Advanced Validation
- What is this?
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RANBP2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 211 bp insertion in exon 1 and 82 bp insertion in exon 1.
View Alternative Names
358 kDa nucleoporin, ANE1, E3 SUMO-protein ligase RanBP2, IIAE3, NUP358, Nuclear pore complex protein Nup358, Nucleoporin 358, Nucleoporin Nup358, P270, RANBP2, RBP2_HUMAN, Ran-binding protein 2, TRP 2, Transformation related protein 2
- WB
Lab
Western blot - Human RANBP2 knockout HeLa cell line (AB265618)
Lanes 1 - 4 : Merged signal (red and green). Green - ab64276 observed at 450 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab64276 was shown to react with RanBP2 in wild-type HeLa cells in Western blot with loss of signal observed in RANBP2 knockout cell line ab265618 (RANBP2 knockout cell lysate ab257627). Wild-type HeLa and RANBP2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab64276 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 : 1000 dilution and 1 : 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 : 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-RanBP2 antibody (<a href='/en-us/products/primary-antibodies/ranbp2-antibody-ab64276'>ab64276</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
RANBP2 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human RANBP2 knockout HeLa cell line (ab265618)
Lane 3:
Caco2 cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
Predicted band size: 358 kDa
Observed band size: 450 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RANBP2 knockout HeLa cell line (AB265618)
Allele-1 : 82 bp insertion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human RANBP2 knockout HeLa cell line (AB265618)
Allele-2 : 211 bp insertion in exon 1.
Reactivity data
Product details
Recommended control: Human wild-type HeLa cell line (ab255928). 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.
We will provide viable cells that proliferate on revival.
This 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.
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
RanBP2 plays a significant role in multiple processes such as mitotic spindle assembly and chromatin condensation. It functions as part of the Ran GTPase system a critical complex for nucleocytoplasmic transport. In addition RanBP2 contributes to protein sumoylation where it acts as a scaffold for conjugating enzymes. It maintains nuclear envelope integrity and supports the positioning of centrosomes during cell division.
Pathways
RanBP2 is actively involved in the nucleocytoplasmic transport pathway ensuring the proper exchange of proteins and RNA between the nucleus and cytoplasm. It is critical to the Ran cycle where it works alongside proteins like RanGTP and RanGAP1 to regulate transport activities. The involvement of RanBP2 in the SUMOylation pathway also emphasizes its role in post-translational modification impacting various cellular events and signaling.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
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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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