AB277860

Human CXCL10 (IP10) knockout THP-1 cell line

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CXCL10 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: 80 bp Deletion in Exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

10 kDa interferon gamma-induced protein, C X C motif chemokine 10, C7, CXCL10, CXCL10(1-73), CXL10_HUMAN, Chemokine (C X C motif) ligand 10, Chemokine CXC motif ligand 10, Crg 2, Gamma-IP10, IFI10, INP 10, Interferon activated gene 10, Interferon gamma induced cell line, Interferon gamma induced factor MOB1, mouse, homolog of, Interferon gamma induced protein 10, Interferon inducible cytokine IP 10, Mob 1, Protein 10 from interferon (gamma) induced cell line, SCYB10, Small inducible cytokine B10 precursor, Small inducible cytokine subfamily B (Cys X Cys) member 10, Small inducible cytokine subfamily B CXC member 10, Small inducible cytokine subfamily B, member 10, Small-inducible cytokine B10, gIP 10

3 Images

Lab

Western blot - Human CXCL10 (IP10) knockout THP-1 cell line (AB277860)

False colour image of Western blot : Anti-IP10 antibody [EPR7850] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab137018 was shown to bind specifically to IP10. A band was observed at 11 kDa in treated wild-type THP-1 cell lysates with no signal observed at this size in treated CXCL10 knockout cell line ab277860 (knockout cell lysate ab283141). To generate this image wild-type and CXCL10 knockout THP-1 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°. 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 (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-IP10 antibody [EPR7850] (<a href='/products/primary-antibodies/ip10-antibody-epr7850-ab137018'>ab137018</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 vehicle control IFNg (0 ng/ml, 32 h), TNF-alpha (0 ng/ml, 32 h), Brefeldin A (5 ug/ml, 6 h) cell lysate at 20 µg

Lane 2:

Wild-type THP-1 treated IFNg (100 ng/ml, 32 h), TNF-alpha (10 ng/ml, 32 h), Brefeldin A (5 ug/ml, 6 h) cell lysate at 20 µg

Lane 3:

CXCL10 knockout THP-1 vehicle control IFNg (0 ng/ml, 32 h), TNF-alpha (0 ng/ml, 32 h), Brefeldin A (5 ug/ml, 6 h) cell lysate at 20 µg

Lane 4:

CXCL10 knockout THP-1 treated IFN-gamma (100 ng/ml, 32 h), TNF-alpha (10 ng/ml, 32 h), Brefeldin A (5 ug/ml, 6 h) cell lysate at 20 µg

Predicted band size: 10 kDa

Observed band size: 11 kDa

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Lab

Western blot - Human CXCL10 (IP10) knockout THP-1 cell line (AB277860)

Lanes 1 - 6 : Merged signal (red and green). Green - ab214668 observed at 11 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab214668 was shown to react with IP10 in wild-type A549 cells in western blot with loss of signal observed in IP10 knockout cell line ab266971 (knockout cell lysate ab256888). Wild-type and IP10 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab214668 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° 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^® 800CW) preabsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-IP10 antibody [EPR20764] (<a href='/products/primary-antibodies/ip10-antibody-epr20764-ab214668'>ab214668</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 6h) cell lysate at 30 µg

Lane 2:

Wild-type A549 IFN-y (<a href='/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) (100 ng/ml, 32 h) and TNF-alpha (<a href='/products/proteins-peptides/recombinant-human-tnf-alpha-protein-active-ab259410'>ab259410</a>) (10 ng/ml, 32h), and Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml for the last 6h) cell lysate at 30 µg

Lane 2:

Western blot - Human CXCL10 (IP10) knockout THP-1 cell line (ab277860)

Lane 3:

IP10 knockout A549 Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 6h) cell lysate at 30 µg

Lane 4:

IP10 knockout A549 IFN-y (<a href='/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) (100ng/ml, 32h) and TNF-alpha (<a href='/products/proteins-peptides/recombinant-human-tnf-alpha-protein-active-ab259410'>ab259410</a>) (10ng/ml, 32h), and Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml for the last 6h) cell lysate at 30 µg

Lane 5:

THP-1 Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 6h) cell lysate at 30 µg

Lane 6:

THP-1 IFN-y (<a href='/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) (200ng/ml, 24h) and LPS (50ng/ml, 24h)-treated for 24 hours, and Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml for the last 6h) cell lysate at 30 µg

Predicted band size: 10 kDa

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Sanger Sequencing - Human CXCL10 (IP10) knockout THP-1 cell line (AB277860)

Sanger seq

Lab

Sanger Sequencing - Human CXCL10 (IP10) knockout THP-1 cell line (AB277860)

80 bp Deletion in Exon 2

Key facts

細胞タイプ

THP-1

生物種

Human

組織

Blood

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, Homozygous: 80 bp Deletion in Exon 2

疾病

Acute Monocytic Leukemia

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Reactivity data

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製品の詳細

We will provide viable cells that proliferate on revival.

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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製品内容

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出荷温度及び保存条件

遺伝子名

CXCL10

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

接合型

Homozygous

出荷温度

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

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取り扱い方法

初回取り扱いガイドライン

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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x10⁵ cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO₂. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x10⁵ cells/mL.Small amounts of fresh media can be added until cell number/viability improves.

継代培養ガイドライン

All seeding densities should be based on cell counts gained by established methods.
Cells should be seeded at 2x10⁵ - 3x10⁵ cells/mL and subcultured when they have reached 8x10⁵ cells/mL.
It is not recommended to allow the cell density to exceed 1x10⁶ cells/mL.

培養培地

RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol

凍結保存培地

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

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補足情報

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

IP10 also known as CXCL10 is a small cytokine belonging to the CXC chemokine family. It has a molecular mass of approximately 8.7 kDa. IP10 is secreted by several cell types such as monocytes endothelial cells and fibroblasts in response to interferon-gamma (IFN-γ). This protein is involved in immune responses and exhibits various roles especially in chemoattracting cells. Researchers often measure IP10 concentrations using ELISA kits such as the IP-10 ELISA to study its expression levels in different biological contexts.

Biological function summary

IP10 plays a role in modulating the activities of immune cells. It attracts T cells eosinophils monocytes and natural killer (NK) cells by binding to the CXCR3 receptor. IP10 is not part of a larger complex but interacts with other cytokines to influence cell migration and the immune response. High levels of IP10 can reflect strong immune activation which is why it is often measured in inflammatory conditions using standard assays like the IP-10 ELISA kits.

Pathways

The role of IP10 lies within the Th1-type immune response pathway. In this pathway IP10 works alongside other chemokines to recruit and activate immune cells to sites of inflammation or infection. It synergizes with IFN-γ to propagate immune signals. IP10 is also linked with the CXCR3 receptor which plays a critical role in these pathways providing a connection to other proteins such as CXCL9 and CXCL11 which have similar functions in cell-mediated immunity.

IP10 is associated with conditions like multiple sclerosis and rheumatoid arthritis. Elevated IP10 levels often correlate with disease activity in these disorders making it a potential biomarker for disease progression. The protein interacts with other inflammatory mediators such as TNF-α in regulating immune activity within these disease contexts. IP10's involvement in recruiting immune cells contributes to the pathogenic inflammation observed in these conditions.

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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

Suspension

Gender

Male

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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

See full target information CXCL10

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Human CXCL10 (IP10) knockout THP-1 cell line