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AB258020

Human LAMTOR2 knockout HeLa cell lysate

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LAMTOR2 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2 and 7 bp deletion in exon2.

View Alternative Names

ENDAP, Endosomal adaptor protein p14, HSPC003, LTOR2_HUMAN, Late endosomal/lysosomal Mp1-interacting protein, Late endosomal/lysosomal adaptor and MAPK and MTOR activator 2, MAPBPIP, MAPKSP1 adaptor protein, MAPKSP1AP, Mitogen-activated protein-binding protein-interacting protein, P14, ROBLD 3, RP11 336K24.9, Ragulator complex protein LAMTOR2, Ragulator2, Roadblock domain containing 3, Roadblock domain-containing protein 3

3 Images
Western blot - Human LAMTOR2 knockout HeLa cell lysate (AB258020)
  • WB

Lab

Western blot - Human LAMTOR2 knockout HeLa cell lysate (AB258020)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : LAMTOR2 knockout HeLa cell lysate (20 μg)

Lane 3 : HepG2 cell lysate (20 μg)

Lane 4 : LNCaP cell lysate (20 μg)

Lanes 1-4 : Merged signal (red and green). Green - ab183514 observed at 14 kDa. Red - loading control ab8245 observed at 37 kDa.

ab183514 Anti-LAMTOR2 antibody [EPR14378] was shown to specifically react with LAMTOR2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265828 (knockout cell lysate ab258020) was used. Wild-type and LAMTOR2 knockout samples were subjected to SDS-PAGE. ab183514 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-LAMTOR2 antibody [EPR14378] (<a href='/en-us/products/primary-antibodies/lamtor2-antibody-epr14378-ab183514'>ab183514</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

LAMTOR2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human LAMTOR2 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-lamtor2-knockout-hela-cell-line-ab265828'>ab265828</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

LNCaP 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 1/20000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

Sanger Sequencing - Human LAMTOR2 knockout HeLa cell lysate (AB258020)
  • Sanger seq

Unknown

Sanger Sequencing - Human LAMTOR2 knockout HeLa cell lysate (AB258020)

Allele-2 : 1 bp deletion in exon2

Sanger Sequencing - Human LAMTOR2 knockout HeLa cell lysate (AB258020)
  • Sanger seq

Unknown

Sanger Sequencing - Human LAMTOR2 knockout HeLa cell lysate (AB258020)

Allele-1 : 7 bp deletion in exon2

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2 and 7 bp deletion in exon2.

Disease

Adenocarcinoma

Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

What's included?

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Properties and storage information

Gene name
LAMTOR2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C

Supplementary information

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

LAMTOR2 also called p14 acts as an important component of the Ragulator complex. With a molecular mass of approximately 16 kDa LAMTOR2 localizes mainly on the late endosomal and lysosomal membranes. This protein aids in the recruitment of other proteins to these membranes facilitating various cellular processes. The expression of LAMTOR2 finds prominence in many tissues especially those with high metabolic activity.
Biological function summary

LAMTOR2 serves in the regulation of mTORC1 signaling which plays a major role in cell growth and metabolism. As a member of the Ragulator complex LAMTOR2 works in concert with other proteins like LAMTOR1 and LAMTOR3 to anchor the Rag GTPases to the lysosomal surface. This provides a platform for the activation of mTORC1 in response to amino acid availability therefore guiding cellular energy and protein synthesis pathways.

Pathways

LAMTOR2 functions integrally in the amino acid-sensing pathway and impacts the mTOR signaling pathway. The Ragulator complex involving LAMTOR2 interacts with Rag GTPases to activate mTORC1 upon amino acid availability bridging nutrient sensing and cellular metabolism control. In doing so it interacts closely with proteins like RagA/RagB coordinating signals that are critical for growth-related cellular functions.

LAMTOR2 disruptions have links to immunodeficiency and some metabolic conditions. Deficient Ragulator function including LAMTOR2 leads to improper mTOR signaling impacting immune cell function and systemic metabolism. Notably this situation associates with diseases like Charcot-Marie-Tooth disease where LAMTOR2's interaction with LAMTOR3 becomes significant due to affected lysosomal positioning and signaling pathways.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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For full details, please see our Terms & Conditions

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