Human REL knockout HCT116 cell line
- Advanced Validation
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REL KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
Avian reticuloendotheliosis, C REL, C Rel protein, Oncogene REL, Oncogene REL avian reticuloendotheliosis, Proto-oncogene c-Rel, REL_HUMAN, V rel reticuloendotheliosis viral oncogene homolog (avian), c Rel proto oncogene protein, v rel avian reticuloendotheliosis viral oncogene homolog, v rel reticuloendotheliosis viral oncogene homolog
- NGS
Lab
Next Generation Sequencing - Human REL knockout HCT116 cell line (AB286471)
48 bp deletion (allele 1), and 48 bp deletion with 2 bp insertion (allele 2) after Gln15 of the WT protein
- WB
Lab
Western blot - Human REL knockout HCT116 cell line (AB286471)
Western blot : Anti-c-Rel antibody [EPR2559(2)] ab133251 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 74 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in REL knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-c-Rel antibody [EPR2559(2)] (<a href='/en-us/products/primary-antibodies/c-rel-antibody-epr25592-ab133251'>ab133251</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 at 20 µg
Lane 2:
Western blot - Human REL knockout HCT116 cell line (ab286471) at 20 µg
Lane 3:
HeLa at 20 µg
Lane 4:
PC-3 at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 69 kDa
Observed band size: 74 kDa,37 kDa
false
Reactivity data
Product details
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.
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
McCoY5a + 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
C-Rel functions to regulate immune system processes and is integral in lymphocyte activation. c-Rel functions as part of both homodimer and heterodimer complexes often partnering with other members of the NF-kappaB family to exert its effects. Through these complexes c-Rel influences the transcription of cytokines growth factors and other molecules critical for immune function. The protein’s role extends to influencing the differentiation and survival of lymphocytes instrumental in mounting adequate immune responses.
Pathways
C-Rel plays a significant role in the regulation of the NF-kappaB signaling pathway and the MAPK (mitogen-activated protein kinase) pathway. The NF-kappaB pathway is activated by inflammatory stimuli and stress which leads to the translocation of c-Rel to the nucleus. Here c-Rel regulates genes important for immune and inflammatory responses alongside other proteins like RelA and p50. In conjunction with the MAPK pathway c-Rel contributes to cellular responses to growth signals and stress collaborating with proteins such as JNK (c-Jun N-terminal kinase) and ERK (extracellular signal-regulated kinase).
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
![Immunoprecipitation - Anti-c-Rel antibody [EPR2559(2)] (AB133251)](https://content.abcam.com/products/images/c-rel-antibody-epr25592-ab133251--immunoprecipitation-img6110.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com