AB301019

Human ITGB4 knockout A549 cell line

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ITGB4 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout. 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

CD 104, CD104 antigen, ITB4_HUMAN, ITG B4, Integrin beta 4 subunit, Integrin beta-4, gp150

3 Images

Lab

Western blot - Human ITGB4 knockout A549 cell line (AB301019)

Western blot : Anti-ITGB4 antibody [EPR17517] (ab182120) 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, ab182120 was shown to bind specifically to ITGB4. A band was observed at 200 kDa in wild-type A549 cell lysates with no signal observed at this size in ITGB4 knockout cell line. To generate this image, wild-type and ITGB4 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-Integrin beta 4 antibody [EPR17517] (<a href='/products/primary-antibodies/integrin-beta-4-antibody-epr17517-ab182120'>ab182120</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 40 µg

Lane 2:

ITGB4 knockout A549 cell lysate at 40 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: 200 kDa

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Lab

Western blot - Human ITGB4 knockout A549 cell line (AB301019)

Western blot : Anti-ITGB4 antibody [EPR8558(2)] (ab168386) staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab168386 was shown to bind specifically to ITGB4. A band was observed at 200 kDa in wild-type A549 cell lysates with no signal observed at this size in ITGB4 knockout cell line. To generate this image, wild-type and ITGB4 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-Integrin beta 4 antibody [EPR8558(2)] (<a href='/products/primary-antibodies/integrin-beta-4-antibody-epr85582-ab168386'>ab168386</a>) at 1/2000 dilution

Lane 1:

Wild-type A549 cell lysate at 40 µg

Lane 2:

ITGB4 knockout A549 cell lysate at 40 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: 200 kDa

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Lab

Next Generation Sequencing - Human ITGB4 knockout A549 cell line (AB301019)

14 bp deletion after His 267 and 131 bp deletion after Val 282 (allele 1); 139 bp deletion after His 267 (allele 2); 9 bp deletion and 1 bp insertion after His 267 and 2 bp deletion after Cys 284 (allele 3); 3 bp insertion after His 267 (allele 4)

Key facts

細胞タイプ

A549

生物種

Human

組織

Lung

製品の状態

Liquid

form

ノックアウト検証方法

Next Generation Sequencing,Western blot

ノックアウト変異

Knockout

疾病

Carcinoma

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Reactivity data

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

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製品の詳細

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.

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.

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製品内容

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab301019-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab301019 Human ITGB4 knockout A549 cell line", "number":"AB301019-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab288558 Human wild-type A549 cell line", "number":"AB301019-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab301019-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab301019 Human ITGB4 knockout A549 cell line", "number":"AB301019-CMP01", "productcode":"" } ] } } }

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出荷温度及び保存条件

遺伝子名

ITGB4

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

Next Generation 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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取り扱い方法

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.
Do not allow the cell density to exceed 7x10⁴ cells/cm².

培養培地

F-12K + 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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補足情報

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

Integrin beta 4 also known as ITGB4 integrin Я4 or integrine beta 4 is a transmembrane protein that mechanically links the extracellular matrix to the intracellular cytoskeleton. This protein has a molecular mass of approximately 205 kDa. Integrin beta 4 primarily expresses in epithelial cells particularly those forming stratified and transitional epithelia. It partners with alpha 6 integrin to form the alpha 6 beta 4 integrin complex which plays a significant role in the formation and maintenance of hemidesmosomes.

Biological function summary

Integrin beta 4 is important for anchoring epithelial cells to the basement membrane stabilizing cell layers in tissues. It interacts with intermediate filaments through specific cytoplasmic proteins. The integrin beta 4 complex ensures structural integrity essential for maintaining the skin and mucosal barriers. Integrations with proteins like M126 are essential for strengthening these interactions safeguarding cellular stability and promoting wound healing.

Pathways

Integrin beta 4 is embedded in key signaling pathways that influence cell adhesion and migration. It participates prominently in the PI3K/AKT and RAS pathway collaborations facilitating signal transduction essential for cell growth and survival. Through these pathways integrin beta 4 interacts with proteins like IS200BPIRG1A linking extracellular cues to internal cellular responses.

Integrin beta 4 has associations with epidermolysis bullosa and various cancers. In epidermolysis bullosa mutations in integrin beta 4 impair hemidesmosomal function leading to skin fragility. In cancer integrin beta 4 overexpression or mislocalization contributes to tumor progression affecting cell invasion and metastasis. Its interactions with IS200BPIAG1A or related pathways highlight the protein’s influence on carcinogenic processes and potential as a therapeutic target.

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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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 ITGB4

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Human ITGB4 knockout A549 cell line