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AB306652

Human DLG4 knockout U-87 MG cell line

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DLG4 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'.
4 Images
Western blot - Human DLG4 knockout U-87 MG cell line (AB306652)
  • WB

Lab

Western blot - Human DLG4 knockout U-87 MG cell line (AB306652)

Western blot : Anti-DLG4 antibody [EPR23124-118] (ab238135) staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab238135 was shown to bind specifically to DLG4. A band was observed at 80 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in DLG4 knockout cell line. To generate this image, wild-type and DLG4 knockout U-87 MG 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-PSD95 antibody [EPR23124-118] - Synaptic Marker (<a href='/en-us/products/primary-antibodies/psd95-antibody-epr23124-118-synaptic-marker-ab238135'>ab238135</a>) at 1/2000 dilution

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lane 2:

Western blot - Human DLG4 knockout U-87 MG cell line (ab306652)

Lane 2:

DLG4 knockout U-87 MG cell lysate at 20 µg

Lane 3:

Wild-type SH-SY5Y cell lysate at 20 µg

Lane 4:

DLG4 knockout SH-SY5Y <a href='/en-us/products/cell-lines/human-psd95-knockout-sh-sy5y-cell-line-ab280043'>ab280043</a> cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

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Western blot - Human DLG4 knockout U-87 MG cell line (AB306652)
  • WB

Lab

Western blot - Human DLG4 knockout U-87 MG cell line (AB306652)

Western blot : Anti-DLG4 antibody (ab18258) staining at 1 ug/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab18258 was shown to bind specifically to DLG4. A band was observed at 80 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in DLG4 knockout cell line. To generate this image, wild-type and DLG4 knockout U-87 MG 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-PSD95 antibody - Synaptic Marker (<a href='/en-us/products/primary-antibodies/psd95-antibody-synaptic-marker-ab18258'>ab18258</a>) at 1 µg/mL

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lane 2:

DLG4 knockout U-87 MG cell lysate at 20 µg

Lane 3:

Wild-type SH-SY5Y ab275475 cell lysate at 20 µg

Lane 4:

DLG4 knockout SH-SY5Y <a href='/en-us/products/cell-lines/human-psd95-knockout-sh-sy5y-cell-line-ab280043'>ab280043</a> cell lysate at 20 µg

Lane 5:

Human Brain cell lysate at 20 µg

Lane 6:

HL-60 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: 80 kDa

false

Western blot - Human DLG4 knockout U-87 MG cell line (AB306652)
  • WB

Lab

Western blot - Human DLG4 knockout U-87 MG cell line (AB306652)

Western blot : Anti-DLG4 antibody (ab18258) staining at 1 ug/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab18258 was shown to bind specifically to DLG4. A band was observed at 80 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in DLG4 knockout cell line. To generate this image, wild-type and DLG4 knockout U-87 MG 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-PSD95 antibody - Synaptic Marker (<a href='/en-us/products/primary-antibodies/psd95-antibody-synaptic-marker-ab18258'>ab18258</a>) at 1 µg/mL

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lane 2:

DLG4 knockout U-87 MG cell lysate at 20 µg

Lane 3:

Wild-type SH-SY5Y ab275475 cell lysate at 20 µg

Lane 4:

DLG4 knockout SH-SY5Y <a href='/en-us/products/cell-lines/human-psd95-knockout-sh-sy5y-cell-line-ab280043'>ab280043</a> cell lysate at 20 µg

Lane 5:

Human Brain cell lysate at 20 µg

Lane 6:

HL-60 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: 80 kDa

false

Next Generation Sequencing - Human DLG4 knockout U-87 MG cell line (AB306652)
  • NGS

Supplier Data

Next Generation Sequencing - Human DLG4 knockout U-87 MG cell line (AB306652)

1 bp insertion and 69 bp deletion after Val124 of the WT protein

Key facts

Cell type

U-87 MG

Species or organism

Human

Tissue

Brain

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout.

Disease

Glioblastoma

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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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Product details

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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What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab306652-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab279979 Human wild-type U-87 MG cell line", "number":"AB306652-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab306652 Human DLG4 knockout U-87 MG cell line", "number":"AB306652-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab306652-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab306652 Human DLG4 knockout U-87 MG cell line", "number":"AB306652-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
DLG4
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
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.
Culture medium

EMEM + 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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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 DLG4
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Product promise

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