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AB258252

Human TRMT1 (TRM1) knockout HEK-293T cell lysate

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TRMT1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon 6.
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Western blot - Human TRMT1 (TRM1) knockout HEK-293T cell lysate (AB258252)
  • WB

Lab

Western blot - Human TRMT1 (TRM1) knockout HEK-293T cell lysate (AB258252)

Lane 1 : Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate, 20 ug

Lane 2 : TRMT1 (TRM1) knockout HEK293T (ab266705) whole cell lysate, 20 ug

Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate, 20 ug

Lanes 1-3 : Merged signal (red and green). Green - ab283652 observed at 75 kDa. Red - loading control ab8245 observed at 36 kDa. ab283652 Anti-TRMT1 (TRM1) antibody [EPR25054-72] was shown to specifically react with TRMT1 (TRM1) in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266705 (knockout cell lysate ab258252) was used. Wild-type and TRMT1 (TRM1) knockout samples were subjected to SDS-PAGE. ab283652 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 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 1/10000 dilution for 1 hour at room temperature before imaging.

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

The bands nearby 100 kDa could be non-specific bands.

All lanes:

Western blot - Anti-TRM1 antibody [EPR25054-72] (<a href='/en-us/products/primary-antibodies/trm1-antibody-epr25054-72-ab283652'>ab283652</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

TRMT1 (TRM1) knockout HEK293T (<a href='/en-us/products/cell-lines/human-trmt1-trm1-knockout-hek-293t-cell-line-ab266705'>ab266705</a>) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TRMT1 (TRM1) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-trmt1-trm1-knockout-hek-293t-cell-line-ab266705'>ab266705</a>)

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>), 1/10000 dilution

Predicted band size: 72 kDa

Observed band size: 75 kDa

false

Western blot - Human TRMT1 (TRM1) knockout HEK-293T cell lysate (AB258252)
  • WB

Lab

Western blot - Human TRMT1 (TRM1) knockout HEK-293T cell lysate (AB258252)

Lane 1 : Wild-type HEK-293T cell lysate 20 μg
Lane 2 : TRMT1 knockout HEK-293T cell lysate 20 μg
Lane 3 : HeLa cell lysate 20 μg
Lane 4 : HepG2 cell lysate 20 μg
False colour image of Western blot : Anti-TRM1 antibody - C-terminal staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab185997 was shown to bind specifically to TRM1. A band was observed at 75 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in Trmt1 knockout cell line ab266705 (knockout cell lysate ab258252). To generate this image, wild-type and Trmt1 knockout HEK-293T 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-TRM1 antibody - C-terminal (<a href='/en-us/products/primary-antibodies/trm1-antibody-c-terminal-ab185997'>ab185997</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

TRMT1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human TRMT1 (TRM1) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-trmt1-trm1-knockout-hek-293t-cell-line-ab266705'>ab266705</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Predicted band size: 72 kDa

Observed band size: 75 kDa

false

Sanger Sequencing - Human TRMT1 (TRM1) knockout HEK-293T cell lysate (AB258252)
  • Sanger seq

Unknown

Sanger Sequencing - Human TRMT1 (TRM1) knockout HEK-293T cell lysate (AB258252)

Allele-2 : Insertion of the selection cassette in exon 6

Sanger Sequencing - Human TRMT1 (TRM1) knockout HEK-293T cell lysate (AB258252)
  • Sanger seq

Unknown

Sanger Sequencing - Human TRMT1 (TRM1) knockout HEK-293T cell lysate (AB258252)

Allele-1 : Insertion of the selection cassette in exon 6

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon 6.

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Complementary Products

Primary Antibodies

AB283652

Anti-TRM1 antibody [EPR25054-72]

20ul selling size|RabMAb|Recombinant|KO Validated|Trial Size|Trial Size

Western blot - Anti-TRM1 antibody [EPR25054-72] (AB283652)

  • 4
  • 1
Primary Antibodies

AB14824

Anti-PPM1A antibody [p6c7]

KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPM1A antibody [p6c7] (AB14824)

  • 5
  • 3
Cell Lines & Lysates

AB266705

Human TRMT1 (TRM1) knockout HEK-293T cell line

Advanced Validation

Western blot - Human TRMT1 (TRM1) knockout HEK-293T cell line (AB266705)

  • 0
  • 0
Primary Antibodies

AB171978

Anti-TIMMDC1/C3orf1 antibody [EPR11807]

RabMAb|Recombinant

Immunocytochemistry/ Immunofluorescence - Anti-TIMMDC1/C3orf1 antibody [EPR11807] (AB171978)

  • 0
  • 0
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Reactivity data

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

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab258252-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human TRMT1 knockout HEK293T cell lysate", "number":"AB258252-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB258252-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
TRMT1
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
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Supplementary information

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

TRM1 also known as transfer RNA methyltransferase 1 is an enzyme responsible for methylating the N2 position of guanine in tRNA molecules. It has a molecular mass of approximately 49 kDa. TRM1 mostly resides in the mitochondria but can also be found in the cytoplasm. Its presence ensures proper methylation of tRNA influencing efficient and accurate protein translation.
Biological function summary

The methylation activity of TRM1 plays a role in maintaining the stability and function of tRNA molecules. As part of the cellular machinery TRM1 does not function alone; it often works together with other tRNA-modifying enzymes to ensure the fidelity of protein synthesis. This modification protects tRNA's structure facilitates proper folding and enhances interaction with ribosomal components.

Pathways

TRM1 functions significantly impact protein synthesis pathways. The enzyme is instrumental in the translation process by ensuring that tRNA molecules are correctly modified for protein assembly. TRM1 operates alongside other proteins such as TRM10 and TRMT5 within these modification pathways influencing the efficiency and accuracy of translation. It supports processes like tRNA maturation and ribosomal activity.

Alterations in TRM1 activity can relate to mitochondrial pathologies. Mutations or deficiencies in TRM1 can disrupt mitochondrial function leading to disorders such as mitochondrial myopathy. Research also connects TRM1 aberrations to neurodegenerative diseases where it may interact with proteins like PINK1 highlighting its role in neuroprotective responses. The efficiency of TRM1 in performing its tasks is vital as any dysfunction can result in profound cellular consequences affecting human health.
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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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Product protocols

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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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