AB267063

Human TRIM56 knockout A549 cell line

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TRIM56 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 3 and 1 bp deletion in exon 3 and 31 bp deletion in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

A130009K11Rik, DKFZp667O116, E3 ubiquitin-protein ligase TRIM56, FLJ35608, Gm452, MGC37358, OTTMUSP00000027392, RING finger protein 109, RNF109, TRI56_HUMAN, Tripartite motif-containing protein 56

6 Images

Lab

Western blot - Human TRIM56 knockout A549 cell line (AB267063)

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 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 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='/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 (ab267063)

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='/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

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Supplier Data

Western blot - Human TRIM56 knockout A549 cell line (AB267063)

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 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 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='/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 (ab267063)

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='/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 Sequencing - Human TRIM56 knockout A549 cell line (AB267063)

Sanger seq

Unknown

Sanger Sequencing - Human TRIM56 knockout A549 cell line (AB267063)

Allele-3 : 1 bp deletion in exon 3.

Sanger seq

Unknown

Sanger Sequencing - Human TRIM56 knockout A549 cell line (AB267063)

Allele-2 : 13 bp deletion in exon 3.

Cell Culture - Human TRIM56 knockout A549 cell line (AB267063)

Cell Culture

Lab

Cell Culture - Human TRIM56 knockout A549 cell line (AB267063)

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.

Sanger seq

Unknown

Sanger Sequencing - Human TRIM56 knockout A549 cell line (AB267063)

Allele-1 : 31 bp deletion in exon3

Key facts

細胞タイプ

A549

生物種

Human

組織

Lung

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 3 and 1 bp deletion in exon 3 and 31 bp deletion in exon 3

疾病

Carcinoma

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Reactivity data

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製品の詳細

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.

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製品内容

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出荷温度及び保存条件

遺伝子名

TRIM56

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

出荷温度

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

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取り扱い方法

初回取り扱いガイドライン

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.

継代培養ガイドライン

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.

培養培地

F-12K + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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補足情報

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

TRIM56 also known as Tripartite Motif Containing 56 or RNF109 is a protein involved in various cellular mechanisms. It weighs around 79 kDa and is expressed in many tissues including the spleen lymph nodes and lungs. TRIM56 contains a RING finger domain B-box motifs and a coiled-coil region which are typical features of the TRIM protein family. These domains suggest roles in ubiquitination and protein-protein interactions which are essential for its functions in signaling and regulatory processes.

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.

TRIM56 exhibits significant connections to viral infections and autoimmune diseases. Its role in antiviral responses implicates it in diseases like hepatitis C as it can suppress viral replication. Although TRIM56 aids in defending against infections its dysregulation may also link to autoimmune conditions where its interaction with proteins such as IFNAR1 can result in excessive immune responses leading to tissue damage and inflammation. Such dual roles highlight TRIM56 as an important regulator in both protective and pathological immune responses.

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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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製品プロトコール

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

See full target information TRIM56

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Human TRIM56 knockout A549 cell line