Human CBLB knockout HeLa cell line
- Advanced Validation
- What is this?
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CBLB KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 6.
View Alternative Names
AI429560, AI851073, CBLB_HUMAN, Cas Br M (murine) ecotropic retroviral transforming sequence b, Casitas B lineage lymphoma b, Casitas B-lineage lymphoma proto-oncogene b, DKFZp686J10223, DKFZp779A0729, DKFZp779F1443, E3 ubiquitin-protein ligase CBL-B, EC 6.3.2.-, FLJ36865, FLJ41152, Nbla00127, RING finger protein 56, RNF56, SH3-binding protein CBL-B, Signal transduction protein CBL-B
- WB
Lab
Western blot - Human CBLB knockout HeLa cell line (AB264711)
Lanes 1-4 : Merged signal (red and green). Green - ab54362 observed at 130 kDa. Red - loading control, ab181602 observed at 37 kDa.
ab54362 Anti-CBLB antibody [246C5a] was shown to specifically react with CBLB in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264711 (knockout cell lysate ab258342) was used. Wild-type and CBLB knockout samples were subjected to SDS-PAGE. ab54362 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-CBLB antibody [246C5a] (<a href='/en-us/products/primary-antibodies/cblb-antibody-246c5a-ab54362'>ab54362</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
CBLB knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human CBLB knockout HeLa cell line (ab264711)
Lane 3:
CTLL-2 cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
Predicted band size: 109 kDa
Observed band size: 130 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CBLB knockout HeLa cell line (AB264711)
Homozygous : 1 bp deletion in exon 6.
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 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
Zygosity
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
CBLB plays a critical role in immune regulation and is part of the ubiquitin-proteasome system. As a member of the CBL protein family it interacts with several other proteins to form a complex that determines its function. This complex mediates signal transduction and maintains immune homeostasis efforting its regulatory influence over immune cell activation and avoiding excessive immune responses.
Pathways
CBLB integrates into intracellular signaling involving the T-cell receptor (TCR) and B-cell receptor (BCR) pathways. It modulates downstream signaling by finetuning protein levels and activity interfacing with related proteins such as ZAP70 and Syk. By controlling signal thresholds it ensures proper immune responses and prevents autoimmunity maintaining the balance in signal transduction processes.
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
Complementary Products
Primary Antibodies
AB315022
Anti-CBLB antibody [EPR28082-32] - BSA and Azide free
RabMAb|KO Validated|Recombinant
![Western blot - Anti-CBLB antibody [EPR28082-32] - BSA and Azide free (AB315022)](https://content.abcam.com/products/images/cblb-antibody-epr28082-32-bsa-and-azide-free-ab315022--western-blot-img423737.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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