AB257429

Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate

Name: Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate (ab257429) | Abcam
Brand: Abcam Limited
SKU: AB257429

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EPHB4 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.

3 Images

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Western blot - Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate (AB257429)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : EPHB4 knockout HEK293T cell lysate (20 ug)
Lane 3 : MCF7 cell lysate (20 ug)
Lane 4 : HT-29 cell lysate (20 ug)

ab254300 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 ab257429) was used. Wild-type and Eph receptor B4/HTK knockout samples were subjected to SDS-PAGE. ab254300 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^oC at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/eph-receptor-b4-htk-antibody-epr23221-54-ab254300'>ab254300</a>) 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 (<a href='/en-us/products/cell-lines/human-ephb4-eph-receptor-b4-htk-knockout-hek-293t-cell-line-ab266733'>ab266733</a>)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

HT-29 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 108 kDa

Observed band size: 125 kDa

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Lab

Western blot - Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate (AB257429)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : EPHB4 knockout HEK293T cell lysate (20 ug)
Lane 3 : MCF7 cell lysate (20 ug)
Lane 4 : HT-29 cell lysate (20 ug)

ab254301 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 ab257429) was used. Wild-type and Eph receptor B4/HTK knockout samples were subjected to SDS-PAGE. ab254301 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^oC at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/eph-receptor-b4-htk-antibody-epr23222-24-ab254301'>ab254301</a>) 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 (<a href='/en-us/products/cell-lines/human-ephb4-eph-receptor-b4-htk-knockout-hek-293t-cell-line-ab266733'>ab266733</a>)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

HT-29 cell lysate at 20 µg

Secondary

All lanes:

Predicted band size: 108 kDa

Observed band size: 125 kDa

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Sanger Sequencing - Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate (AB257429)

Sanger seq

Unknown

Sanger Sequencing - Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate (AB257429)

Homozygous : 1 bp insertion in exon3

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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What's included?

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Properties and storage information

Gene name

EPHB4

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Shipped at conditions

Ambient - Can Ship with Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

Biological function summary

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.

Pathways

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.

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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate

Western blot - Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate (AB257429)

Western blot - Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate (AB257429)

Sanger Sequencing - Human EPHB4 (Eph receptor B4/HTK) knockout HEK-293T cell lysate (AB257429)

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Complementary Products

Primary Antibodies

Proteins & Peptides

Proteins & Peptides

Proteins & Peptides

Reactivity data

Product details

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

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Product promise