Human CXCL10 (IP10) knockout A549 cell lysate (ab256888)
Overview
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Product name
Human CXCL10 (IP10) knockout A549 cell lysate
See all IP10 kits -
Product overview
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) -
Parental Cell Line
A549 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and 4 bp deletion in exon2. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
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 - found here. 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.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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.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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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab263518 - Human CXCL10 knockout A549 cell lysate - Brefeldin A treated 1 x 100µg ab277648 - Human CXCL10 knockout A549 cell lysate - IFN-y, TNF-alpha + Brefeldin A treated 1 x 100µg ab277649 - Human wild-type A549 cell lysate - Brefeldin A treated 1 x 100µg ab277650 - Human wild-type A549 cell lysate - IFN-y, TNF-alpha + Brefeldin A treated 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Carcinoma -
STR Analysis
Amelogenin X,Y D5S818: 11 D13S317: 11 D7S820: 8, 11 D16S539: 11, 12 vWA: 14 TH01: 8,9.3 TPOX: 8,11 CSF1PO: 10, 12
Target
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Function
Chemotactic for monocytes and T-lymphocytes. Binds to CXCR3. -
Sequence similarities
Belongs to the intercrine alpha (chemokine CxC) family. -
Post-translational
modificationsCXCL10(1-73) is produced by proteolytic cleavage after secretion from keratinocytes. -
Cellular localization
Secreted. - Information by UniProt
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Alternative names
- Interferon gamma induced factor MOB1, mouse, homolog of
- Interferon gamma induced protein 10
- 10 kDa interferon gamma induced protein
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab256888 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 10 kDa.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 10 kDa. |
Images
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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.
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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 ugBlocking 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.
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Allele-1: 4 bp deletion in exon2
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Allele-2: 1 bp insertion in exon2
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab256888 has not yet been referenced specifically in any publications.