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AB319092

Human DUSP4 knockout A549 cell line [H15]

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DUSP4 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 109 bp deletion (allele 1) and 62 bp deletion (allele 2) in exon 2, CCDS6072.1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
3 Images
Western blot - Human DUSP4 knockout A549 cell line [H15] (AB319092)
  • WB

Lab

Western blot - Human DUSP4 knockout A549 cell line [H15] (AB319092)

Western blot : Anti-DUSP4 antibody [EPR19881] (ab216576) 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, ab216576 was shown to bind specifically to DUSP4. A band was observed at 40 kDa in wild-type A549 cell lysates with no signal observed at this size in DUSP4 knockout cell line. To generate this image, wild-type and DUSP4 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-DUSP4 antibody [EPR19881] (<a href='/en-us/products/primary-antibodies/dusp4-antibody-epr19881-ab216576'>ab216576</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lanes 2 - 3:

DUSP4 knockout A549 cell lysate at 20 µg

Lane 4:

DUSP4 knockout A549 H15 cell lysate at 20 µg

Lane 5:

HCT 116 cell lysate at 20 µg

Lane 6:

MOLT-4 cell lysate at 20 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: 40 kDa

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Western blot - Human DUSP4 knockout A549 cell line [H15] (AB319092)
  • WB

Lab

Western blot - Human DUSP4 knockout A549 cell line [H15] (AB319092)

Western blot : Anti-DUSP4 antibody [EPR19881] (ab216576) 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, ab216576 was shown to bind specifically to DUSP4. A band was observed at 40 kDa in wild-type A549 cell lysates with no signal observed at this size in DUSP4 knockout cell line. To generate this image, wild-type and DUSP4 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.

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

DUSP4 knockout A549 F7 cell lysate at 20 µg

Lane 3:

DUSP4 knockout A549 D4 cell lysate at 20 µg

Lane 4:

DUSP4 knockout A549 H15 cell lysate at 20 µg

Lane 5:

HCT 116 cell lysate at 20 µg

Lane 6:

MOLT-4 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

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Next Generation Sequencing - Human DUSP4 knockout A549 cell line [H15] (AB319092)
  • NGS

Lab

Next Generation Sequencing - Human DUSP4 knockout A549 cell line [H15] (AB319092)

109 bp deletion (allele 1) and 62 bp deletion (allele 2) in exon 2, CCDS6072.1

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, 109 bp deletion (allele 1) and 62 bp deletion (allele 2) in exon 2, CCDS6072.1

Disease

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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What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab319092-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab288558 Human wild-type A549 cell line", "number":"AB319092-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab319092 Human DUSP4 knockout A549 cell line [H15]", "number":"AB319092-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab319092-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab319092 Human DUSP4 knockout A549 cell line [H15]", "number":"AB319092-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
DUSP4
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage duration
A few days
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

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.

Subculture guidelines
  • 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.
Culture medium

F-12K + 10% FBS

Cryopreservation medium

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

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Quality control

STR analysis

D3S1358, TH01, D21S11, D18S51, D5S818, D13S317, D7S820, D16S539, CSF1PO, D8S1179, TPOX, FGA, D19S433, D2S1338, VWA

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Target data

See full target information DUSP4
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Complementary Products

Primary Antibodies

AB216576

Anti-DUSP4 antibody [EPR19881]

RabMAb|Recombinant|KO Validated|20ul selling size

Immunocytochemistry/ Immunofluorescence - Anti-DUSP4 antibody [EPR19881] (AB216576)

  • 0
  • 0
Proteins & Peptides

AB132331

Recombinant Human DUSP4 protein

SDS-PAGE - Recombinant Human DUSP4 protein (AB132331)

  • 0
  • 0

Product promise

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For full details, please see our Terms & Conditions

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