TRIM21 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 4 and 1 bp insertion in exon 4 and 8 bp deletion in exon 4.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 1 bp insertion in exon 4 and 8 bp deletion in exon 4
Alternative names
52 kDa Ro protein, 52 kDa ribonucleoprotein autoantigen Ro/SS-A, 52kD Ro/SSA autoantigen, Autoantigen Ro/SSA, 52-KD, E3 ubiquitin-protein ligase TRIM21, RING finger protein 81, RNF81, RO52_HUMAN, Ro(SS-A), SS-A, SSA1: Sjogren syndrome antigen A1 (52kDa ribonucleoprotein autoantigen SS-A/Ro), Sicca syndrome antigen A, Sjoegren syndrome type A antigen, Sjogren syndrome antigen A1, Sjogren syndrome type A antigen, TRIM21, Tripartite motif protein TRIM21, Tripartite motif-containing 21, Tripartite motif-containing protein 21
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TRIM21 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 4 and 1 bp insertion in exon 4 and 8 bp deletion in exon 4.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 1 bp insertion in exon 4 and 8 bp deletion in exon 4
- Antibiotic resistance
- Puromycin 1µg/mL
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- TRIM21
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
We will provide viable cells that proliferate on revival.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
TRIM21 also known as SS-A or Ro52 is a 52 kDa protein that plays a role in the immune system. It functions as an E3 ubiquitin ligase involved in the ubiquitination process. This protein is expressed in various tissues including immune cells like lymphocytes and macrophages. TRIM21 recognizes autoantibodies linked to TRIM21 activity particularly those targeting the SS-A complex. Researchers often use antibodies like anti-TRIM21 or anti-SS-A to study its interactions and effects in cellular processes.
- Biological function summary
TRIM21 contributes significantly to the regulation of immune responses and participates in innate and adaptive immunity. It forms part of a complex with other proteins to facilitate the degradation of viral particles through the ubiquitin-proteasome system a process known as antibody-dependent intracellular neutralization. As an important player in the immune defense TRIM21 oversees the timely removal of pathogens and prevents potential overactivation of immune responses that might harm the host.
- Pathways
TRIM21 operates within the interferon signaling and NF-κB pathways two important areas of immune response modulation. TRIM21 interacts with molecules like transcription factors that influence the expression of interferon-responsive genes which are critical for pathogen defense. Its role in these pathways highlights its interactions with various immune-regulatory proteins helping to maintain immune system balance and effectiveness during infections.
- Associated diseases and disorders
TRIM21 has strong associations with autoimmune conditions such as systemic lupus erythematosus and Sjögren's syndrome. Autoantibodies against TRIM21 are often used as biomarkers for these diseases and their presence can indicate increased autoimmunity. Relationship with other autoantigens like SS-A/Ro60 further connects TRIM21 with disease mechanisms suggesting its importance in the pathology of these autoimmune disorders. Understanding the role of TRIM21 in these diseases may improve diagnostic and therapeutic strategies.
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5 product images
Western blot - Human TRIM21 (SS-A) knockout A549 cell line (ab267024)
Lanes 1-4: Merged signal (red and green). Green - Anti-TRIM21/SS-A antibody [EPR20290] ab207728 observed at 50 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.Anti-TRIM21/SS-A antibody [EPR20290] ab207728 Anti-TRIM21/SS-A antibody [EPR20290] was shown to specifically react with TRIM21/SS-A in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267024 (knockout cell lysate Human TRIM21 (SS-A) knockout A549 cell lysate ab257766) was used. Wild-type and TRIM21/SS-A knockout samples were subjected to SDS-PAGE. Anti-TRIM21/SS-A antibody [EPR20290] ab207728 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-TRIM21/SS-A antibody [EPR20290] (Anti-TRIM21/SS-A antibody [EPR20290] ab207728) at 1/1000 dilution
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: TRIM21 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human TRIM21 (SS-A) knockout A549 cell line (ab267024)
Lane 3: HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 4: MOLT-4 (Human lymphoblastic leukemia cell line) whole cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 54 kDa
Observed band size: 50 kDa
Cell Culture - Human TRIM21 (SS-A) knockout A549 cell line (ab267024)
Representative images TRIM21 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.
Sanger Sequencing - Human TRIM21 (SS-A) knockout A549 cell line (ab267024)
Allele-2: 1 bp deletion in exon 4.
Sanger Sequencing - Human TRIM21 (SS-A) knockout A549 cell line (ab267024)
Allele-1: 8 bp deletion in exon4
Sanger Sequencing - Human TRIM21 (SS-A) knockout A549 cell line (ab267024)
Allele-3: 1 bp insertion in exon 4.
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