CXCL10 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9,
Homozygous: 80 bp Deletion in Exon 2.
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
CXCL10 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9,
Homozygous: 80 bp Deletion in Exon 2.
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.
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.
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.
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.
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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False colour image of Western blot: Anti-IP10 antibody [EPR7850] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-IP10 antibody [EPR7850] 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-IP10 antibody [EPR7850] (Anti-IP10 antibody [EPR7850] ab137018) 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 2: Western blot - Human CXCL10 (IP10) knockout THP-1 cell line (ab277860)
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
Performed under reducing conditions.
Predicted band size: 10 kDa
Observed band size: 11 kDa
Anti-IP10 antibody [EPR20764] 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 Human CXCL10 (IP10) knockout A549 cell line ab266971 (knockout cell lysate Human CXCL10 (IP10) knockout A549 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 Anti-IP10 antibody [EPR20764] ab214668 and Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-IP10 antibody [EPR20764] (Anti-IP10 antibody [EPR20764] ab214668) at 1/1000 dilution
Lane 1: Wild-type A549 Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299)-treated (5ug/ml, 6h) cell lysate at 30 µg
Lane 2: Wild-type A549 IFN-y (Recombinant Human Interferon gamma protein (Active) ab259377) (100 ng/ml, 32 h) and TNF-alpha (Recombinant human TNF alpha protein (Active) ab259410) (10 ng/ml, 32h), and Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299)-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 (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299)-treated (5ug/ml, 6h) cell lysate at 30 µg
Lane 4: IP10 knockout A549 IFN-y (Recombinant Human Interferon gamma protein (Active) ab259377) (100ng/ml, 32h) and TNF-alpha (Recombinant human TNF alpha protein (Active) ab259410) (10ng/ml, 32h), and Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299)-treated (5ug/ml for the last 6h) cell lysate at 30 µg
Lane 5: THP-1 Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299)-treated (5ug/ml, 6h) cell lysate at 30 µg
Lane 6: THP-1 IFN-y (Recombinant Human Interferon gamma protein (Active) ab259377) (200ng/ml, 24h) and LPS (50ng/ml, 24h)-treated for 24 hours, and Brefeldin A (Brefeldin A, Inhibitor of ADP-ribosylation factor ab120299)-treated (5ug/ml for the last 6h) cell lysate at 30 µg
Performed under reducing conditions.
Predicted band size: 10 kDa
80 bp Deletion in Exon 2
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