EPHB4 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 insertion in exon 3.
EPHB4_HUMAN, Ephrin receptor EphB4, Ephrin type-B receptor 4, HTK, Hepatoma Transmembrane Kinase, MYK 1, TYRO 11, Tyrosine-protein kinase TYRO11, Tyrosine-protein kinase receptor HTK
EPHB4 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 insertion in exon 3.
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.
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.
The Eph receptor B4 also known as EphB4 or HTK is a protein belonging to the Ephrin receptor subfamily of receptor tyrosine kinases. The receptor has a molecular weight of about 120 kDa. EphB4 typically interacts with its binding partner ephrin-B2 facilitating bidirectional signaling. These complexes reside in the cell membrane and are mainly found in vascular endothelial cells and various tissues including the heart lungs and skin. Notably this receptor plays a role in cellular processes like cell migration and adhesion.
The Eph receptor B4 is a significant regulator of angiogenesis and vascular development. It forms a critical component of a signaling complex that also includes ephrin-B2. During embryonic development EphB4's interaction with ephrin-B2 guides the proper formation and organization of blood vessels. In adult organisms it remains active in maintaining vascular homeostasis. Additionally this receptor impacts neuronal development and repair through modulating axon guidance and synaptic plasticity.
EphB4 engages in various cellular signaling cascades. It features prominently within pathways governing angiogenesis and cell adhesion. EphB4 acts as a central player in the VEGF signaling pathway modulating endothelial cell behavior. The protein's pathways frequently intersect with other proteins such as FAK and Src influencing cell motility and morphology. Furthermore in the ephrin receptor pathway it cross-talks with other Eph receptors like EphA4 assisting in diverse biological functions.
Abnormal Eph receptor B4 activity associates with several pathological conditions. Dysregulation often links to cancer due to its role in neovascularization and tumor progression. Overexpression of EphB4 in tumors like breast and colorectal cancer suggests a contribution to malignancy by supporting angiogenesis and tumor growth. In vascular disorders impaired EphB4-ephrin-B2 signaling may lead to defects in blood vessel formation. Additionally alterations in this receptor are observed in neurological diseases such as Alzheimer's where EphB4 interacts with tau protein-modifying vascular and neuronal deficiencies.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
Anti-Eph receptor B4/HTK antibody [EPR23221-54] ab254300 Anti-Eph receptor B4/HTK antibody [EPR23221-54] was shown to specifically react with Eph receptor B4/HTK in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266733 (knockout cell lysate Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate ab257429) was used. Wild-type and Eph receptor B4/HTK knockout samples were subjected to SDS-PAGE. Anti-Eph receptor B4/HTK antibody [EPR23221-54] ab254300 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Eph receptor B4/HTK antibody [EPR23221-54] (Anti-Eph receptor B4/HTK antibody [EPR23221-54] ab254300) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: EPHB4 knockout HEK293T cell lysate at 20 µg
Lane 2: Western blot - Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell line (ab266733)
Lane 3: MCF7 cell lysate at 20 µg
Lane 4: HT-29 cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 108 kDa
Observed band size: 125 kDa
Anti-Eph receptor B4/HTK antibody [EPR23222-24] ab254301 Anti-Eph receptor B4/HTK antibody [EPR23222-24] was shown to specifically react with Eph receptor B4/HTK in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266733 (knockout cell lysate Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate ab257429) was used. Wild-type and Eph receptor B4/HTK knockout samples were subjected to SDS-PAGE. Anti-Eph receptor B4/HTK antibody [EPR23222-24] ab254301 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Eph receptor B4/HTK antibody [EPR23222-24] (Anti-Eph receptor B4/HTK antibody [EPR23222-24] ab254301) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: EPHB4 knockout HEK293T cell lysate at 20 µg
Lane 2: Western blot - Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell line (ab266733)
Lane 3: MCF7 cell lysate at 20 µg
Lane 4: HT-29 cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 108 kDa
Observed band size: 125 kDa
Representative images EPHB4 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
Homozygous: 1 bp insertion in exon3
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