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AB267024

Human TRIM21 (SS-A) knockout A549 cell line

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TRIM21 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. 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. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
5 Images
Western blot - Human TRIM21 (SS-A) knockout A549 cell line (AB267024)
  • WB

Lab

Western blot - Human TRIM21 (SS-A) knockout A549 cell line (AB267024)

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 ab267024 (knockout cell lysate ab257766) 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° 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-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 (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

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

false

Sanger Sequencing - Human TRIM21 (SS-A) knockout A549 cell line (AB267024)
  • Sanger seq

Unknown

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)
  • Sanger seq

Unknown

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)
  • Sanger seq

Unknown

Sanger Sequencing - Human TRIM21 (SS-A) knockout A549 cell line (AB267024)

Allele-3 : 1 bp insertion in exon 4.

Cell Culture - Human TRIM21 (SS-A) knockout A549 cell line (AB267024)
  • Cell Culture

Lab

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.

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

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

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Complementary Products

Primary Antibodies

AB207728

Anti-TRIM21/SS-A antibody [EPR20290]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRIM21/SS-A antibody [EPR20290] (AB207728)

  • 0
  • 0
Primary Antibodies

AB179434

Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448]

20ul selling size|RabMAb|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (AB179434)

  • 5
  • 6
Cell Lines & Lysates

AB257767

Human TRIM21 (SS-A) knockout A549 cell lysate

Western blot - Human TRIM21 (SS-A) knockout A549 cell lysate (AB257767)

  • 0
  • 0
Cell Lines & Lysates

AB306820

Human IDH1 knockout U-87 MG cell line

Next Generation Sequencing - Human IDH1 knockout U-87 MG cell line (AB306820)

  • 5
  • 1
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

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.

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What's included?

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Properties and storage information

Gene name
TRIM21
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Target data

See full target information TRIM21
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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