人CXCL10 (IP10) knockout A549 cell裂解物
Human CXCL10 (IP10) knockout A549 cell lysate
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CXCL10 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and 4 bp deletion in exon2.
查看别名
Gamma-IP10, INP10, C-X-C motif chemokine 10, CXCL10, Small-inducible cytokine B10, SCYB10, 10 kDa interferon gamma-induced protein, IP-10
- WB
Lab
Western blot - Human CXCL10 (IP10) knockout A549 cell lysate (AB256888)
Lane 1 : Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 6h) cell lysate (30 ug)
Lane 2 : Wild-type A549 IFN-y (ab259377) (100 ng/ml, 32 h) and TNF-alpha (ab259410) (10 ng/ml, 32h), and Brefeldin A (ab120299)-treated (5ug/ml for the last 6h) cell lysate (30 ug)
Lane 3 : IP10 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 6h) cell lysate (30 ug)
Lane 4 : IP10 knockout A549 IFN-y (ab259377) (100ng/ml, 32h) and TNF-alpha (ab259410) (10ng/ml, 32h), and Brefeldin A (ab120299)-treated (5ug/ml for the last 6h) cell lysate (30 ug)
Lane 5 : THP-1 Brefeldin A (ab120299)-treated (5ug/ml, 6h) cell lysate (30 ug)
Lane 6 : THP-1 IFN-y (ab259377) (200ng/ml, 24h) and LPS (50ng/ml, 24h)-treated for 24 hours, and Brefeldin A (ab120299)-treated (5ug/ml for the last 6h) cell lysate (30 ug)
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°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® 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 (<a href='/products/cell-lines/human-cxcl10-ip10-knockout-thp-1-cell-line-ab277860'>ab277860</a>)
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
false
- WB
Supplier Data
Western blot - Human CXCL10 (IP10) knockout A549 cell lysate (AB256888)
Lane 1 : Untreated Wild-type A549 (human lung carcinoma epithelial cell), whole cell lysate, 40 ug
Lane 2 : Wild-type A549 treated with 100 ng/ml IFN-y (ab259377) for 32 hours and 10 ng/m TNF-alpha (ab259410) for 32 hours, and 5ug/ml Brefeldin A (ab120299) for the last 6 hours, whole cell lysate, 40 ug
Lane 3 : Untreated IP10 knockout A549 whole cell lysate, 40 ug
Lane 4 : IP10 knockout A549 treated with 100 ng/ml IFN-y (ab259377) for 32 hours and 10 ng/m TNF-alpha (ab259410) for 32 hours, and 5ug/ml Brefeldin A (ab120299) for the last 6 hours, whole cell lysate, 40 ug
Lane 5 : Untreated THP-1 (human monocytic leukemia monocyte), whole cell lysate, 20 ug
Lane 6 : THP-1 treated with 200ng/ml IFN-y (ab259377) for 24 hours and 50ng/ml LPS for 24 hours, and 5ug/ml Brefeldin A for the last 21 hours, whole cell lysate, 20 ug
Blocking and diluting buffer and concentration : 5% NFDM/TBST
Lanes 1-6 : Merged signal (red and green). Green - ab283681 observed at 11 kDa. Red-loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) was observed at 36 kDa.
ab283681 Anti-TNF Receptor I antibody [EPR24674-12] was shown to specifically react with IP10 in treated wild-type A549 cells. Loss of signal was observed when IP10 knockout cell lines ab266971 (knockout cell lysate ab256888) were used. Wild-type and IP10 knockout samples were subjected to SDS-PAGE. ab283681 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-IP10 antibody [EPR24674-12] (<a href='/products/primary-antibodies/ip10-antibody-epr24674-12-ab283681'>ab283681</a>) at 1/1000 dilution
Lane 1:
Untreated Wild-type A549 (human lung carcinoma epithelial cell), whole cell lysate at 40 µg
Lane 2:
Wild-type A549 treated with 100 ng/ml IFN-y (<a href='/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) for 32 hours and 10 ng/m TNF-alpha (<a href='/products/proteins-peptides/recombinant-human-tnf-alpha-protein-active-ab259410'>ab259410</a>) for 32 hours, and 5ug/ml Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) for the last 6 hours, whole cell lysate at 40 µg
Lane 3:
Untreated IP10 knockout A549 whole cell lysate at 40 µg
Lane 4:
IP10 knockout A549 treated with 100 ng/ml IFN-y (<a href='/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) for 32 hours and 10 ng/m TNF-alpha (<a href='/products/proteins-peptides/recombinant-human-tnf-alpha-protein-active-ab259410'>ab259410</a>) for 32 hours, and 5ug/ml Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) for the last 6 hours, whole cell lysate at 40 µg
Lane 5:
Untreated THP-1 (human monocytic leukemia monocyte), whole cell lysate at 20 µg
Lane 6:
THP-1 treated with 200ng/ml IFN-y (<a href='/products/proteins-peptides/recombinant-human-interferon-gamma-protein-active-ab259377'>ab259377</a>) for 24 hours and 50ng/ml LPS for 24 hours, and 5ug/ml Brefeldin A for the last 21 hours, whole cell lysate at 20 µg
Secondary
All lanes:
Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution
Predicted band size: 10 kDa
Observed band size: 11 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CXCL10 (IP10) knockout A549 cell lysate (AB256888)
Allele-2 : 1 bp insertion in exon2
- Sanger seq
Unknown
Sanger Sequencing - Human CXCL10 (IP10) knockout A549 cell lysate (AB256888)
Allele-1 : 4 bp deletion in exon2
搭配使用产品
Primary Antibodies
AB272363
Anti-IP10 antibody [EPR18685-158] - BSA and Azide free (Detector)
RabMAb|Recombinant
![Sandwich ELISA - Anti-IP10 antibody [EPR18685-158] - BSA and Azide free (Detector) (AB272363)](https://content.abcam.com/products/images/ip10-antibody-epr18685-158-bsa-and-azide-free-detector-ab272363--sandwich-elisa-img51812.jpg)
- 0
- 0
反应性数据
产品详情
Knockout cell lysate achieved by CRISPR/Cas9.
Treatments:
Human CXCL10 knockout A549 cell lysate - Brefeldin A (5 μg/ml, 6h)
Human wild-type A549 cell lysate - Brefeldin A (5 μg/ml, 6h)
Human CXCL10 knockout A549 cell lysate - IFN-y (100 ng/ml, 32h), TNF-a (10 ng/ml, 32h) and Brefeldin A (5 μg/ml, during the last 6h of IFN-y/TNF-a treatment)
Human wild-type A549 cell lysate - IFN-y (100 ng/ml, 32h), TNF-a (10 ng/ml, 32h) and Brefeldin A (5 μg/ml, during the last 6h of IFN-y/TNF-a treatment)
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.
规格
性能和储存信息
基因名称
基因编辑类型
基因编辑方法
敲除验证
运输条件
推荐的短期储存条件
推荐的长期储存条件
补充信息
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.
质量控制
STR 分析
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
细胞培养
生物安全等级
EU: 1 US: 1
贴壁/悬浮
Adherent
性别
Male
Abcam Product Promise
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