AB255375

Human CD9 knockout HeLa cell line

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CD9 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

View Alternative Names

5H9, 5H9 antigen, Antigen defined by monoclonal antibody 60229, BA-2/p24 antigen, BA2, BTCC 1, CD9 antigen, CD9 antigen p24, CD9 molecule, CD9_HUMAN, Cell growth-inhibiting gene 2 protein, DRAP 27, GIG2, Growth inhibiting gene 2 protein, Leukocyte antigen MIC3, MIC3, Motility-related protein, Tetraspanin-29, Tspan-29, p24, p24 antigen

4 Images

Lab

Western blot - Human CD9 knockout HeLa cell line (AB255375)

Lanes 1 - 4 : Merged signal (red and green). Green - ab58989 observed at 24 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

ab58989 was shown to react with CD9 in wild-type HeLa cells in Western blot with loss of signal observed in CD9 knockout cell line ab255375 (CD9 knockout cell lysate ab263754). Wild-type HeLa and CD9 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab58989 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-CD9 antibody [TS9] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/cd9-antibody-ts9-bsa-and-azide-free-ab58989'>ab58989</a>) at 1 µg/mL

Lane 1:

Wild-type HeLa (No DTT) cell lysate at 20 µg

Lane 2:

CD9 knockout HeLa (No DTT) cell lysate at 20 µg

Lane 2:

Western blot - Human CD9 knockout HeLa cell line (ab255375)

Lane 3:

A549 (Non-Reducing) cell lysate at 20 µg

Lane 4:

MCF7 (Non-Reducing) cell lysate at 20 µg

Predicted band size: 25 kDa

Observed band size: 24 kDa

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Lab

Western blot - Human CD9 knockout HeLa cell line (AB255375)

Lanes 1 - 4 : Merged signal (red and green). Green - ab263019 observed at 18 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab263019 was shown to react with CD9 in wild-type HeLa cells in Western blot with loss of signal observed in CD9 knockout cell line ab255375 (CD9 knockout cell lysate ab263754). Wild-type HeLa and CD9 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab263019 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-CD9 antibody [EPR23105-125] (<a href='/en-us/products/primary-antibodies/cd9-antibody-epr23105-125-ab263019'>ab263019</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

CD9 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CD9 knockout HeLa cell line (ab255375)

Lane 3:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 25 kDa

Observed band size: 18 kDa

false

Lab

Western blot - Human CD9 knockout HeLa cell line (AB255375)

Lanes 1 - 4 : Merged signal (red and green). Green - ab236630 observed at 18 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab236630 was shown to react with CD9 in wild-type HeLa cells in Western blot with loss of signal observed in CD9 knockout cell line ab255375 (CD9 knockout cell lysate ab263754). Wild-type HeLa and CD9 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab236630 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-CD9 antibody [EPR23105-121] (<a href='/en-us/products/primary-antibodies/cd9-antibody-epr23105-121-ab236630'>ab236630</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

CD9 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CD9 knockout HeLa cell line (ab255375)

Lane 3:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 25 kDa

Observed band size: 18 kDa

false

Sanger Sequencing - Human CD9 knockout HeLa cell line (AB255375)

Sanger seq

Unknown

Sanger Sequencing - Human CD9 knockout HeLa cell line (AB255375)

Homozygous : 1 bp insertion in exon 3.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 3

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "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

CD9

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Zygosity

Homozygous

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

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.

CD9 also known as motility-related protein-1 (MRP-1) is a transmembrane glycoprotein with a molecular weight of approximately 24 kDa. This protein is part of the tetraspanin family and shows widespread expression throughout different tissues including hematopoietic and non-hematopoietic cells. CD9 participates in numerous cellular processes by interacting with integrins and other cell surface receptors facilitating the organization of molecular complexes within the cell membrane.

Biological function summary

The CD9 protein is integral to cell adhesion migration and fusion. It forms complexes with other tetraspanins and proteins like EWI-2 and EWI-F contributing to cellular signaling and membrane compartmentalization. Additionally CD9 plays an important role in the formation and secretion of exosomes tiny vesicles emitted by cells that mediate cell-to-cell communication. These exosomes can be analyzed using tools like exosome assays and detection kits that target CD9 proteins.

Pathways

CD9 is involved in the regulation of immune response and cell morphology. It participates in pathways such as the integrin signaling pathway and the phosphatidylinositol 3-kinase (PI3K) signaling pathway. CD9 modulates these interactions by associating with proteins such as integrins and other members of the tetraspanin family influencing cellular movement and proliferation.

CD9 has connections to various conditions including cancer and infectious diseases. In cancer CD9 can influence tumor progression and metastasis while its altered expression levels have been associated with different cancer types. CD9 interacts with proteins such as integrins and CD81 in these contexts affecting cellular adhesion and migration mechanisms. In infectious diseases CD9 is involved in viral entry processes as some viruses utilize CD9 and its associated tetraspanins for entry into host cells.

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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Target data

See full target information CD9

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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 CD9 knockout HeLa cell line

Western blot - Human CD9 knockout HeLa cell line (AB255375)

Western blot - Human CD9 knockout HeLa cell line (AB255375)

Western blot - Human CD9 knockout HeLa cell line (AB255375)

Sanger Sequencing - Human CD9 knockout HeLa cell line (AB255375)

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

Zygosity

Shipped at conditions

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

Target data

Complementary Products

Primary Antibodies

Proteins & Peptides

Primary Antibodies

Secondary Antibodies

Product promise