AB265178

Human ENG (CD105) knockout HeLa cell line

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ENG KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 2 and 19 bp deletion in exon 2 and 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

END, CD105, Endoglin, ENG

5 Images

Lab

Western blot - Human ENG (CD105) knockout HeLa cell line (AB265178)

ab170943 Anti-CD105 antibody [EPR10145-10] was shown to specifically react with CD105 in HeLa wild type cells. Loss of signal was observed when knockout cell line ab265178 (knockout cell lysate ab256906) was used. Wild type and CD105 knockout samples were subjected to SDS-PAGE. ab170943 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-CD105 antibody [EPR10145-10] (<a href='/products/primary-antibodies/cd105-antibody-epr10145-10-ab170943'>ab170943</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ENG knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ENG (CD105) knockout HeLa cell line (ab265178)

Lane 3:

HUVEC cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 70 kDa

Observed band size: 70-120 kDa

false

Lab

Western blot - Human ENG (CD105) knockout HeLa cell line (AB265178)

ab169545 Anti-CD105 antibody [EPR10145-12] was shown to specifically react with CD105 in HeLa wild type cells. Loss of signal was observed when knockout cell line ab265178 (knockout cell lysate ab256906) was used. Wild type and CD105 knockout samples were subjected to SDS-PAGE. ab169545 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-CD105 antibody [EPR10145-12] (<a href='/products/primary-antibodies/cd105-antibody-epr10145-12-ab169545'>ab169545</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ENG knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ENG (CD105) knockout HeLa cell line (ab265178)

Lane 3:

HUVEC cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 70 kDa

Observed band size: 70-120 kDa

false

Sanger Sequencing - Human ENG (CD105) knockout HeLa cell line (AB265178)

Sanger seq

Unknown

Sanger Sequencing - Human ENG (CD105) knockout HeLa cell line (AB265178)

Allele-2 : 11 bp deletion in exon 2.

Sanger seq

Unknown

Sanger Sequencing - Human ENG (CD105) knockout HeLa cell line (AB265178)

Allele-1 : 19 bp deletion in exon 2.

Sanger seq

Unknown

Sanger Sequencing - Human ENG (CD105) knockout HeLa cell line (AB265178)

Allele-3 : 1 bp insertion in exon 2.

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 2 and 19 bp deletion in exon 2 and 1 bp insertion in exon 2

抗生物質耐性

Puromycin 1µg/mL

疾病

Adenocarcinoma

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Reactivity data

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

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製品の詳細

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.

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製品内容

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出荷温度及び保存条件

遺伝子名

ENG

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

出荷温度

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

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補足情報

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

CD105 also known as endoglin or the CD105 marker is a transmembrane glycoprotein with a molecular weight of approximately 180 kDa. It is a component of the TGF-beta receptor complex and exists in endothelial cells where it is abundantly expressed. Expression of CD105 is higher in proliferating cells particularly in the vasculature. You can also find it in tissues involved in the formation and remodeling of blood vessels such as during angiogenesis.

Biological function summary

Endoglin functions in the regulation of angiogenesis and vascular remodeling. It plays a significant role in mediating cellular responses to TGF-beta signaling influencing endothelial cell proliferation and migration. While not part of a larger structural complex endoglin interacts with receptors and signaling molecules important for vascular development and repair processes. This involvement aids in maintaining endothelial integrity and function under various physiological conditions.

Pathways

CD105 participates in the TGF-beta signaling and angiogenesis pathways. In these pathways it acts in conjunction with other proteins like TGF-beta receptors which play roles in cell differentiation proliferation and apoptosis. The interaction between CD105 and TGF-beta signaling regulates numerous cellular mechanisms impacting angiogenesis and cellular responses to environmental changes.

CD105 has links to hereditary hemorrhagic telangiectasia (HHT) and certain cancers. In HHT mutations in the endoglin gene alter vascular structure leading to the formation of abnormal blood vessels. Oncologically overexpression of CD105 is present in tumor angiogenesis aiding in the progression of certain cancers. Other proteins like VEGF and TGF-beta closely interact with endoglin influencing disease progression and presenting potential targets for therapeutic intervention.

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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

初回取り扱いガイドライン

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.

継代培養ガイドライン

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.

培養培地

DMEM (High Glucose) + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

See full target information ENG

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Abcam product promise

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Human ENG (CD105) knockout HeLa cell line