AB269613

Human ACSL4 (FACL4) knockout A-431 cell line

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ACSL4 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 26 bp deletion Frameshift: 100%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

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Supplier Data

Next Generation Sequencing - Human ACSL4 (FACL4) knockout A-431 cell line (AB269613)

Knockout achieved by CRISPR/Cas9; X = 26 bp deletion; Frameshift : 100%

Lab

Western blot - Human ACSL4 (FACL4) knockout A-431 cell line (AB269613)

Western blot : Anti-FACL4 antibody [EPR17587] ab205197 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 79 kDa in Wild-type A431 cell lysates with no signal observed at this size in ACSL4 knockout A431 cell line. To generate this image, 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-FACL4 antibody [EPR17587] (<a href='/products/primary-antibodies/facl4-antibody-epr17587-ab205197'>ab205197</a>) at 1/1000 dilution

Lane 1:

Wild-type A431 cell lysates at 20 µg

Lane 2:

Western blot - Human ACSL4 (FACL4) knockout A-431 cell line (ab269613) at 20 µg

Lane 3:

HEK-293 cell lysates at 20 µg

Lane 4:

K562 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 79 kDa

Observed band size: 79 kDa

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Key facts

細胞タイプ

A-431

生物種

Human

組織

Skin

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 achieved by CRISPR/Cas9 X = 26 bp deletion Frameshift: 100%

疾病

Epidermoid Carcinoma

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Reactivity data

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

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出荷温度及び保存条件

遺伝子名

ACSL4

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

短期保存期間

1-2 weeks

短期保存温度

-196°C|-80°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.

FACL4 also known as ACSL4 is a member of the long-chain acyl-CoA synthetase (ACSL) family. This enzyme catalyzes the activation of long-chain fatty acids by forming a thioester bond with coenzyme A. FACL4 plays a critical role in lipid metabolism. The molecular weight of FACL4 is approximately 80 kDa. This protein is expressed in various tissues with a higher presence in the brain and testis reflecting its important function in lipid-related processes in these areas.

Biological function summary

FACL4 contributes significantly to lipid metabolism and is involved in the synthesis of complex lipids. It facilitates the integration of long-chain fatty acids into phospholipids and triglycerides. FACL4 often functions as part of larger lipid synthesis and modification complexes working closely with other enzymes to maintain membrane integrity and signal transduction. Its activity influences cellular responses to changes in lipid availability and metabolic demands.

Pathways

FACL4 functions within the arachidonic acid metabolism and glycerophospholipid metabolic pathways. In these pathways other proteins such as COX and LOX interact with FACL4 to regulate inflammatory responses and membrane lipid composition. The enzyme's role in these pathways highlights its contribution to managing the balance between pro-inflammatory and anti-inflammatory metabolites which are essential for cell signaling and homeostasis.

FACL4 is implicated in several neuropsychiatric conditions including X-linked adrenoleukodystrophy and breast cancer. In the context of these diseases FACL4 interacts with proteins like ALDP in adrenoleukodystrophy. Disrupted FACL4 activity can lead to abnormal lipid accumulation or impaired lipid signaling underlying the pathophysiology of these conditions. Understanding the role of FACL4 in these disorders highlights its potential as a therapeutic target for correcting lipid metabolism dysfunction.

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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

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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Human ACSL4 (FACL4) knockout A-431 cell line