Human TRIM56 knockout A549 cell line
- Advanced Validation
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TRIM56 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 2 bp deletion in exon 3.
View Alternative Names
A130009K11Rik, DKFZp667O116, E3 ubiquitin-protein ligase TRIM56, FLJ35608, Gm452, MGC37358, OTTMUSP00000027392, RING finger protein 109, RNF109, TRI56_HUMAN, Tripartite motif-containing protein 56
- WB
Lab
Western blot - Human TRIM56 knockout A549 cell line (AB267062)
Lanes 1-4 : Merged signal (red and green). Green - ab154821 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.
ab154821 Anti-TRIM56 antibody [EPR10582] was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267062 (knockout cell lysate ab258249) was used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154821 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-TRIM56 antibody [EPR10582] (<a href='/en-us/products/primary-antibodies/trim56-antibody-epr10582-ab154821'>ab154821</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human TRIM56 knockout A549 cell line (ab267062)
Lane 3:
MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 4:
A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 81 kDa
Observed band size: 88 kDa
false
- WB
Lab
Western blot - Human TRIM56 knockout A549 cell line (AB267062)
Lanes 1-4 : Merged signal (red and green). Green - ab154862 observed at 88 kDa. Red - loading control ab8245 observed at 36 kDa.
ab154862 Anti-TRIM56 antibody [EPR10583] was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267062 (knockout cell lysate ab258249) was used. Wild-type and TRIM56 knockout samples were subjected to SDS-PAGE. ab154862 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-TRIM56 antibody [EPR10583] (<a href='/en-us/products/primary-antibodies/trim56-antibody-epr10583-ab154862'>ab154862</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human TRIM56 knockout A549 cell line (ab267062)
Lane 3:
MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 4:
A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 81 kDa
Observed band size: 88 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human TRIM56 knockout A549 cell line (AB267062)
Allele-1 : 2 bp deletion in exon3
- Sanger seq
Unknown
Sanger Sequencing - Human TRIM56 knockout A549 cell line (AB267062)
Allele-2 : 1 bp deletion in exon 3.
- Cell Culture
Lab
Cell Culture - Human TRIM56 knockout A549 cell line (AB267062)
Representative images TRIM56 knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
Reactivity data
Product details
Recommended control: Human wild-type A549 cell line (ab255450). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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
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 seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 10% FBS
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
TRIM56 participates in immune regulation and antiviral defense. It acts as an interferon-stimulated gene product and does not appear to be part of a larger protein complex. Instead it induces antiviral states by mediating the innate immune response to viral infections. TRIM56 enhances the production of type I interferons by modifying the activation pathways. This function places TRIM56 as an immune defense agent which can restrict the replication of various viruses such as flaviviruses and coronaviruses therefore harboring significant implications in pathogen defense responses.
Pathways
TRIM56 integrates into the antiviral and innate immunity pathways. It prominently situates within the interferon signaling pathway enhancing the body's defense through the upregulation of interferon production. TRIM56 interacts with several proteins in these pathways including STING and MAVS which are known to be important mediators in the recognition of viral presence. Through these interactions TRIM56 facilitates the activation of downstream signaling cascades leading to the expression of interferon-stimulated genes.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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