Human EPHB2 knockout HCT116 cell line
- Advanced Validation
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EPHB2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
View Alternative Names
DRT, Developmentally regulated EPH related tyrosine kinase, EK5, ELK related protein tyrosine kinase, EPH tyrosine kinase 3, EPH-like kinase 5, EPHB2_HUMAN, EPHT 3, ERK, ETECK, Eph receptor B2, Ephrin type-B receptor 2, Hek 5, Nuk, Prkm 5, Receptor protein tyrosine kinase HEK 5, Renal carcinoma antigen NY-REN-47, Sek 3, Tyro 5, Tyrosine protein kinase receptor CEK 5, Tyrosine protein kinase receptor QEK 5, Tyrosine-protein kinase TYRO5, Tyrosine-protein kinase receptor EPH-3, cek5
- WB
Lab
Western blot - Human EPHB2 knockout HCT116 cell line (AB287395)
Western blot : Anti-EPHB2 antibody [EPR22422-67] (ab307811) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab307811 was shown to bind specifically to EPHB2. A band was observed at 130 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in EPHB2 knockout cell line. To generate this image, wild-type and EPHB2 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Eph receptor B2 antibody [EPR22422-67] (<a href='/en-us/products/primary-antibodies/eph-receptor-b2-antibody-epr22422-67-ab307811'>ab307811</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
EPHB2 knockout HCT 116 cell lysate at 20 µg
Lane 3:
A549 cell lysate at 20 µg
Lane 4:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 130 kDa
false
- WB
Lab
Western blot - Human EPHB2 knockout HCT116 cell line (AB287395)
Western blot : Anti-EIF2AK3 antibody (ab65142) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab65142 was shown to bind specifically to EIF2AK3. A band was observed at 145 kDa in wild-type A549 cell lysates with no signal observed at this size in EIF2AK3 knockout cell line. To generate this image, wild-type and EIF2AK3 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Eph receptor B2 antibody [EPR22427-268] (<a href='/en-us/products/primary-antibodies/eph-receptor-b2-antibody-epr22427-268-ab252935'>ab252935</a>) at 1/3000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
EPHB2 knockout HCT 116 cell lysate at 20 µg
Lane 2:
Western blot - Human EPHB2 knockout HCT116 cell line (ab287395)
Lane 3:
A549 cell lysate at 20 µg
Lane 4:
HepG2 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 130 kDa
false
- NGS
Lab
Next Generation Sequencing - Human EPHB2 knockout HCT116 cell line (AB287395)
85 bp deletion after Lys 136 and 7 bp deletion after Ala 155 (allele 1); 68 bp deletion after Arg 163 (allele 2) of WT protein
Reactivity data
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HCT116 cell line (ab288559). 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.
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
Eph receptor B2 functions in cell-cell communication and is involved in neuronal navigation and synaptic plasticity. It forms part of a complex that brings together ligand-receptor pairs on adjacent cells facilitating bidirectional signaling. The receptor's activation influences neuronal circuitry remodeling and helps in the differentiation of numerous cell types. The B2 protein acting as anti B2 modulates various cell signaling mechanisms essential for normal physiological functioning especially in the nervous system where it contributes to the intricate network of the eph protein interactions.
Pathways
These interactions place Eph receptor B2 in significant signaling pathways such as the MAPK/ERK and PI3K/Akt pathways. These play influential roles in cell proliferation survival and migration. The receptor interacts with numerous proteins including AB25AP through these pathways affecting developmental processes and mature cellular functions. The precise regulation of these pathways is necessary for maintaining homeostasis in complex biological systems especially within the context of the nervous system.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Complementary Products
Primary Antibodies
AB252935
Anti-Eph receptor B2 antibody [EPR22427-268]
BOND RX™ Validated|20ul selling size|RabMAb|Advanced Validation|Recombinant|KO Validated
![Multiplex immunohistochemistry - Anti-Eph receptor B2 antibody [EPR22427-268] (AB252935)](https://content.abcam.com/products/images/eph-receptor-b2-antibody-epr22427-268-ab252935--multiplex-immunohistochemistry-img410072.jpg)
- 0
- 0
Primary Antibodies
AB61791
Anti-Eph receptor B1 + Eph receptor B2 (phospho Y594 + Y596) antibody
- 5
- 4
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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