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AB263754

Human CD9 knockout HeLa cell lysate

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CD9 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 3.

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
Western blot - Human CD9 knockout HeLa cell lysate (AB263754)
  • WB

Lab

Western blot - Human CD9 knockout HeLa cell lysate (AB263754)

Lane 1 : Wild-type HeLa (No DTT) cell lysate 20 μg
Lane 2 : CD9 knockout HeLa (No DTT) cell lysate 20 μg
Lane 3 : A549 (Non-Reducing) cell lysate 20 μg
Lane 4 : MCF7 (Non-Reducing) cell lysate 20 μg
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 55kDa.
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 (<a href='/en-us/products/cell-lines/human-cd9-knockout-hela-cell-line-ab255375'>ab255375</a>)

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

false

Western blot - Human CD9 knockout HeLa cell lysate (AB263754)
  • WB

Lab

Western blot - Human CD9 knockout HeLa cell lysate (AB263754)

Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 μg
Lane 2 : CD9 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 μg
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate 20 μg
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 μg
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 55kDa.
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 (<a href='/en-us/products/cell-lines/human-cd9-knockout-hela-cell-line-ab255375'>ab255375</a>)

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

Western blot - Human CD9 knockout HeLa cell lysate (AB263754)
  • WB

Lab

Western blot - Human CD9 knockout HeLa cell lysate (AB263754)

Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 μg
Lane 2 : CD9 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 μg
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate 20 μg
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate 20 μg
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 55kDa.
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 (<a href='/en-us/products/cell-lines/human-cd9-knockout-hela-cell-line-ab255375'>ab255375</a>)

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 lysate (AB263754)
  • Sanger seq

Unknown

Sanger Sequencing - Human CD9 knockout HeLa cell lysate (AB263754)

Homozygous : 1 bp insertion in exon 3

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

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

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab263754-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human CD9 knockout HeLa cell lysate", "number":"AB263754-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB263754-CMP01", "productcode":"" } ] } } }
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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
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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

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