Human TRIM21 (SS-A) knockout A549 cell line
- Advanced Validation
- What is this?
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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, 13 bp deletion in exon 4 and 1 bp insertion in exon 4.
View 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
- WB
Lab
Western blot - Human TRIM21 (SS-A) knockout A549 cell line (AB267025)
Lanes 1-4 : Merged signal (red and green). Green - ab207728 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.
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 ab267025 (knockout cell lysate ab257767) was used. Wild-type and TRIM21/SS-A knockout samples were subjected to SDS-PAGE. ab207728 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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-TRIM21/SS-A antibody [EPR20290] (<a href='/en-us/products/primary-antibodies/trim21-ss-a-antibody-epr20290-ab207728'>ab207728</a>) at 1/500 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
TRIM21 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human TRIM21 (SS-A) knockout A549 cell line (ab267025)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
MOLT-4 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: 54 kDa
Observed band size: 50 kDa
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- WB
Supplier Data
Western blot - Human TRIM21 (SS-A) knockout A549 cell line (AB267025)
ab207728 was shown to react with TRIM21 in wild-type A549 cells in Western blot with loss of signal observed in TRIM21 knockout cell line ab267025. Wild-type A549 and TRIM21 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab207728 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes:
Western blot - Anti-TRIM21/SS-A antibody [EPR20290] (<a href='/en-us/products/primary-antibodies/trim21-ss-a-antibody-epr20290-ab207728'>ab207728</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 lysate at 30 µg
Lane 2:
TRIM21 knock-out A549 lysate at 30 µg
false
- Cell Culture
Unknown
Cell Culture - Human TRIM21 (SS-A) knockout A549 cell line (AB267025)
Representative images of 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 an EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human TRIM21 (SS-A) knockout A549 cell line (AB267025)
Allele-1 : 13 bp deletion in exon4
- Sanger seq
Unknown
Sanger Sequencing - Human TRIM21 (SS-A) knockout A549 cell line (AB267025)
Allele-2 : 1 bp insertion in exon 4.
- Sanger seq
Lab
Sanger Sequencing - Human TRIM21 (SS-A) knockout A549 cell line (AB267025)
Sequencing chromatogram displaying sequence edit in exon 4
- sELISA
Supplier Data
Sandwich ELISA - Human TRIM21 (SS-A) knockout A549 cell line (AB267025)
Interpolated concentrations of native TRIM21/SS-A in human control wild type A549 (human lung carcinoma cell) and TRIM21 (TRIM21/SS-A) knockout A549 cell based on 200 µg/mL extract loads. The concentrations of TRIM21/SS-A were measured in duplicate and interpolated from the TRIM21/SS-A standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TRIM21/SS-A concentration was determined to be 7026.2 pg/mL in wild type A549 extract (Human wild-type A549 cell line ab255450) and undetectable in TRIM21 (TRIM21/SS-A) knockout A549 extract (Human TRIM21 (TRIM21/SS-A) knockout A549 cell line ab267025).
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, 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.
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
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.
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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Primary Antibodies
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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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