Human TRIM56 knockout A549 cell lysate (ab258250)
Overview
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Product name
Human TRIM56 knockout A549 cell lysate
See all TRIM56 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
A549 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon3 and 1 bp deletion in exon3 and 31 bp deletion in exon3. -
Passage number
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.
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 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.
Tested applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab263626 - Human TRIM56 knockout A549 cell lysate 1 x 100µg ab255554 - Human wild-type A549 cell lysate 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
E3 ubiquitin-protein ligase that mediates 'Lys-63'-linked polyubiquitination of TMEM173/STING, thereby playing a key role in innate immunity. TMEM173/STING 'Lys-63'-linked ubiquitination activates the production of type I interferon IFN-beta following detection of pathogen- and host-derived double-stranded DNA. -
Sequence similarities
Belongs to the TRIM/RBCC family.
Contains 2 B box-type zinc fingers.
Contains 1 RING-type zinc finger. -
Cellular localization
Cytoplasm. - Information by UniProt
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Alternative names
- A130009K11Rik
- DKFZp667O116
- E3 ubiquitin-protein ligase TRIM56
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab263626 - Human TRIM56 knockout A549 cell lysate 1 x 100µg ab255554 - Human wild-type A549 cell lysate 1 x 100µg -
Research areas
-
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
Images
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Western blot - Human TRIM56 knockout A549 cell lysate (ab258250)
Lane 1:Wild-type A549 (Human lung carcinoma cell line) whole cell lysate (20 ug)
Lane 2:TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate (20 ug)
Lane 3:MCF7 (Human breast adenocarcinoma cell line) whole cell lysate (20 ug)
Lane 4:A-375 (Human malignant melanoma cell line) whole cell lysate (20 ug)ab154821 was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267063 (knockout cell lysate ab258250) 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 4oC 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.
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Western blot - Human TRIM56 knockout A549 cell lysate (ab258250)Lane 1:Wild-type A549 (Human lung carcinoma cell line) whole cell lysate (20 ug)
Lane 2:TRIM56 knockout A549 (Human lung carcinoma cell line) whole cell lysate (20 ug)
Lane 3:MCF7 (Human breast adenocarcinoma cell line) whole cell lysate (20 ug)
Lane 4:A-375 (Human malignant melanoma cell line) whole cell lysate (20 ug)ab154862 was shown to specifically react with TRIM56 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267063 (knockout cell lysate ab258250) 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 4oC 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.
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Sanger Sequencing - Human TRIM56 knockout A549 cell lysate (ab258250)Allele-1: 31 bp deletion in exon3
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Sanger Sequencing - Human TRIM56 knockout A549 cell lysate (ab258250)Allele-2: 13 bp deletion in exon3
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Sanger Sequencing - Human TRIM56 knockout A549 cell lysate (ab258250)Allele-3: 1 bp deletion in exon3
