AB273857

Human MAGEA4 knockout A-431 cell line

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MAGEA4 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 = 2 bp deletion, 1 bp insertion; Frameshift: 99%. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

CT1.4, Cancer/testis antigen 1.4, MAGA4_HUMAN, MAGE 4, MAGE 41, MAGE 4A, MAGE 4B, MAGE X2, MAGE-4 antigen, MAGE-41 antigen, MAGE-X2 antigen, MGC21336, Melanoma antigen family A 4, Melanoma-associated antigen 4

3 Images

Lab

Western blot - Human MAGEA4 knockout A-431 cell line (AB273857)

False colour image of Western blot : Anti-MAGEA4 antibody staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab229011 was shown to bind specifically to MAGEA4. A band was observed at 40 kDa in wild-type A431 cell lysates with no signal observed at this size in MAGEA4 knockout cell line ab273857 (knockout cell lysate ab273811). To generate this image, wild-type and MAGEA4 knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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:

SH-SY5Y cell lysate at 1/500 dilution

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

Western blot - Human MAGEA4 knockout A-431 cell lysate (ab273811) at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

Western blot 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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Lab

Western blot - Human MAGEA4 knockout A-431 cell line (AB273857)

False colour image of Western blot : Anti-MAGEA4 antibody [OTI1F9] staining at 1/4000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab139297 was shown to bind specifically to MAGEA4. A band was observed at 40 kDa in wild-type A431 cell lysates with no signal observed at this size in MAGEA4 knockout cell line ab273857 (knockout cell lysate ab273811). To generate this image, wild-type and MAGEA4 knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-MAGEA4 antibody [OTI1F9] (<a href='/en-us/products/primary-antibodies/magea4-antibody-oti1f9-ab139297'>ab139297</a>) at 1/4000 dilution

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

Western blot - Human MAGEA4 knockout A-431 cell lysate (ab273811) at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 35 kDa

Observed band size: 40 kDa

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Supplier Data

Next Generation Sequencing - Human MAGEA4 knockout A-431 cell line (AB273857)

Knockout achieved by CRISPR/Cas9; X = 2 bp deletion, 1 bp insertion; Frameshift : 99%

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 2 bp deletion, 1 bp insertion; Frameshift: 99%

Disease

Epidermoid 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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Product details

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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What's included?

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Properties and storage information

Gene name

MAGEA4

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 duration

1-2 weeks

Appropriate short-term storage conditions

-196°C|-80°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

DMEM (High Glucose) + 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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Supplementary information

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

MAGEA4 also known as MAGE-A4 is part of the melanoma-associated antigen family. This protein is encoded by the MAGEA4 gene and is characterized by a molecular mass of approximately 42 kDa. Its expression is limited to certain tissues including the testis and is often silent in normal somatic tissues. However MAGEA4 is frequently found in various types of tumors making it a target of interest for cancer research and therapy.

Biological function summary

MAGEA4 participates in cellular processes related to cancer progression. Although not part of a multi-protein complex its presence in tumorous tissues suggests a role in modulating cellular mechanisms that favor tumor survival and proliferation. MAGEA4 does not directly interact with other proteins but might affect cellular pathways indirectly contributing to oncogenesis.

Pathways

MAGEA4 influences several cellular mechanisms that are related to cancer development. While it is not a central player in major biological pathways its function relates to processes affecting cell cycle regulation and apoptosis. This connection was observed in studies showing interactions with other cancer-related proteins like p53 suggesting that MAGEA4 can modify responses to apoptotic signals and may interact with or affect the TP53 pathway.

MAGEA4 is prominently linked to various cancers including melanomas and carcinomas such as lung cancer. Its frequent expression in tumor cells but not in normal tissues makes MAGEA4 a target for cancer immunotherapy. Additionally studies have shown that MAGEA4 may interfere with the immune system's ability to detect and destroy cancerous cells partly due to interactions with proteins like CTLA-4 further emphasizing its potential as a therapeutic target in oncology.

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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

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Human MAGEA4 knockout A-431 cell line

Western blot - Human MAGEA4 knockout A-431 cell line (AB273857)

Western blot - Human MAGEA4 knockout A-431 cell line (AB273857)

Next Generation Sequencing - Human MAGEA4 knockout A-431 cell line (AB273857)

Key facts

Cell type

Species or organism

Tissue

Form

Knockout validation

Mutation description

Disease

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Handling procedures

Initial handling guidelines

Subculture guidelines

Culture medium

Cryopreservation medium

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Complementary Products

Cell Lines & Lysates

Primary Antibodies

Cell Lines & Lysates

Primary Antibodies

Product promise