AB273711

Human TMEM106B knockout A549 cell line

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TMEM106B KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 100%. 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

Transmembrane protein 106B, TMEM106B

4 Images

Lab

Western blot - Human TMEM106B knockout A549 cell line (AB273711)

Western blot : Anti-TMEM106B antibody staining at 0.1 µg/ml, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, the antibody was shown to bind specifically to TMEM106B. A band was observed at 45 kDa in wild-type A549 cell lysates with no signal observed at this size in TMEM106B knockout cell line ab273711 (knockout cell lysate ab273769). To generate this image, wild-type and TMEM106B knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1% Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Anti-TMEM106B antibody staining at 0.1 µg/mL

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human TMEM106B knockout A549 cell lysate (<a href='/products/cell-lysates/human-tmem106b-knockout-a549-cell-lysate-ab273769'>ab273769</a>) at 20 µg

Lane 3:

PANC-1 cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

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Lab

Next Generation Sequencing - Human TMEM106B knockout A549 cell line (AB273711)

1 bp insertion after Arg88 of the WT protein

Supplier Data

Next Generation Sequencing - Human TMEM106B knockout A549 cell line (AB273711)

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift : 100%

Immunocytochemistry/ Immunofluorescence - Human TMEM106B knockout A549 cell line (AB273711)

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Lab

Immunocytochemistry/ Immunofluorescence - Human TMEM106B knockout A549 cell line (AB273711)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Wildtype A549 (wildtype human lung carcinoma epithelial cell); TMEM106B KO A549 (TMEM106B knockout A549), ab273711 cells labelling TMEM106B with ab325555 at 1/500 (0.998 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing lysosome staining in wildtype A549 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab25631 Anti-LAMP2 mouse monoclonal antibody - Lysosome Marker was used to counterstain lysosomes at 1/200 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

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

Next Generation Sequencing

ノックアウト変異

Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 100%

疾病

Carcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" } } }

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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, 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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製品内容

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出荷温度及び保存条件

遺伝子名

TMEM106B

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

Next Generation Sequencing

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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補足情報

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

TMEM106B also known as transmembrane protein 106B is a type II single-pass transmembrane protein with a molecular mass of approximately 33 kDa. It is expressed in specific regions of the brain as well as in other tissue types such as the lungs and immune system cells. Its expression levels are especially elevated in microglia and oligodendrocytes which are brain cells that play important roles in immune response and myelination respectively. Structurally TMEM106B contains a transmembrane domain and its precise biochemical function is still under investigation though it is thought to participate in lysosomal and vesicular trafficking.

Biological function summary

TMEM106B is involved in several cellular processes impacting lysosome function and trafficking. It may become part of a protein complex that manages lysosomal positioning and pathway regulation within the cell. TMEM106B also affects cellular homeostasis and protein degradation which suggests a role in maintaining the proper balance of proteins and lipids within lysosomes. Alteration in TMEM106B expression or function can cause changes in lysosomal size and distribution indicating its role in cellular logistics.

Pathways

The TMEM106B protein becomes involved in the endolysosomal pathway and the autophagy-lysosome pathway. These pathways are critical for processing and breakdown of cellular waste and recycling of cellular components. In these pathways TMEM106B interacts with other proteins such as SORT1 (sortilin 1) and GRN (progranulin) both of which have important roles in protein sorting and trafficking. By influencing these pathways TMEM106B helps maintain cellular health and function.

TMEM106B links to neurodegenerative conditions such as frontotemporal lobar degeneration and possibly Alzheimer's disease. Mutations or abnormal expression levels of TMEM106B have been implicated in the pathogenesis and progression of these diseases. In frontotemporal lobar degeneration TMEM106B interacts with proteins like GRN where changes can lead to neurodegeneration. In Alzheimer's disease abnormal lysosomal function tied to TMEM106B may contribute to the accumulation of pathogenic proteins.

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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

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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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 TMEM106B

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Human TMEM106B knockout A549 cell line