Human CXCL10 (IP10) knockout THP-1 cell line
- Advanced Validation
- What is this?
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- WB
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='/en-us/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
false
- WB
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='/en-us/products/primary-antibodies/ip10-antibody-epr20764-ab214668'>ab214668</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 Brefeldin A (<a href='/en-us/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='/en-us/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) (100 ng/ml, 32 h) and TNF-alpha (<a href='/en-us/products/proteins-peptides/recombinant-human-tnf-alpha-protein-active-ab259410'>ab259410</a>) (10 ng/ml, 32h), and Brefeldin A (<a href='/en-us/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='/en-us/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='/en-us/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) (100ng/ml, 32h) and TNF-alpha (<a href='/en-us/products/proteins-peptides/recombinant-human-tnf-alpha-protein-active-ab259410'>ab259410</a>) (10ng/ml, 32h), and Brefeldin A (<a href='/en-us/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='/en-us/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='/en-us/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='/en-us/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
false
- Sanger seq
Lab
Sanger Sequencing - Human CXCL10 (IP10) knockout THP-1 cell line (AB277860)
80 bp Deletion in Exon 2
Complementary Products
Primary Antibodies
AB306587
Anti-CXCL10/IP-10 antibody [EPR24674-84]
BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCL10/IP-10 antibody [EPR24674-84] (AB306587)](https://content.abcam.com/products/images/cxcl10-ip-10-antibody-epr24674-84-ab306587--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img403870.jpg)
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Reactivity data
Product details
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.
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
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.
Subculture guidelines
- 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.
Culture medium
RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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