AB265351

Human LAMTOR1 knockout HeLa cell line

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LAMTOR1 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 1 and 4 bp insertion in exon 1. 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

C11orf59, LTOR1_HUMAN, Late endosomal/lysosomal adaptor and MAPK and MTOR activator 1, Lipid raft adaptor protein p18, PDRO, PP7157, Protein associated with DRMs and endosomes, Ragulator complex protein LAMTOR1, Ragulator1, RhoA activator C11orf59, p18, p27Kip1-releasing factor from RhoA, p27RF-Rho, ragulator complex protein PDRO

3 Images

Lab

Western blot - Human LAMTOR1 knockout HeLa cell line (AB265351)

Lanes 1 - 4 : Merged signal (red and green). Green - ab121157 observed at 18 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab121157 was shown to react with LAMTOR1 in wild-type HeLa cells in Western blot with loss of signal observed in LAMTOR1 knockout cell line ab265351 (LAMTOR1 knockout cell lysate ab258938). Wild-type HeLa and LAMTOR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab121157 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 0.04 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

Lanes 1 - 4:

Western blot - Anti-LAMTOR1 antibody (<a href='/products/primary-antibodies/lamtor1-antibody-ab121157'>ab121157</a>) at 0.04 µg/mL

Lanes 1 - 4:

Western blot - Anti-LAMTOR1 antibody (<a href='/products/primary-antibodies/lamtor1-antibody-ab229760'>ab229760</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

LAMTOR1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human LAMTOR1 knockout HeLa cell line (ab265351)

Lane 3:

PANC-1 cell lysate at 20 µg

Lane 4:

U-2 OS cell lysate at 20 µg

Predicted band size: 18 kDa

Observed band size: 18 kDa

false

Sanger Sequencing - Human LAMTOR1 knockout HeLa cell line (AB265351)

Sanger seq

Unknown

Sanger Sequencing - Human LAMTOR1 knockout HeLa cell line (AB265351)

Allele-2 : 4 bp insertion in exon 1.

Sanger seq

Unknown

Sanger Sequencing - Human LAMTOR1 knockout HeLa cell line (AB265351)

Allele-1 : 1 bp deletion in exon 1.

Key facts

細胞タイプ

HeLa

生物種

Human

組織

Cervix

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, 1 bp deletion in exon 1 and 4 bp insertion in exon 1

疾病

Adenocarcinoma

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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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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

LAMTOR1

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.

培養培地

DMEM (High Glucose) + 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.

LAMTOR1 also known as p18 is a protein with a mass of approximately 18 kDa. It functions mechanically as a scaffold protein located on the late endosome/lysosome membrane. The protein's primary role involves recruiting and stabilizing the other members of the Ragulator complex. LAMTOR1 is expressed in various tissues with significant levels in immune and neuronal cells where it supports cellular growth and immune responses.

Biological function summary

LAMTOR1 acts as a part of the highly-conserved Ragulator complex. The Ragulator complex is vital for the activation of the mechanistic target of rapamycin complex 1 (mTORC1) in response to amino acids. LAMTOR1's interaction with other components of the complex such as LAMTOR2 and LAMTOR3 facilitates the localization and activation of mTORC1 on the lysosomal surface. This activation promotes anabolic processes including protein lipid and nucleotide synthesis.

Pathways

LAMTOR1 plays an important role in the mTOR signaling pathway. This pathway regulates cell growth proliferation and metabolism. LAMTOR1's involvement with Rag proteins assists in sensing amino acid levels thereby influencing the mTORC1 pathway activation. Additionally LAMTOR1-related pathways intersect with the AMPK signaling pathway where it functionally interacts with proteins that control cellular energy homeostasis.

LAMTOR1's dysregulation has been associated with conditions like cancer and neurodegenerative disorders. Altered expression of LAMTOR1 can impact mTORC1 signaling leading to uncontrolled cell growth often observed in cancerous cells. In neurodegenerative disorders defects in mTORC1 pathway can cause neuronal cell damage. These conditions link LAMTOR1 to proteins like Rheb and TSC2 which are important for maintaining a balance in these signaling pathways.

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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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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 LAMTOR1

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Human LAMTOR1 knockout HeLa cell line