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AB258253

Human TRMT2A (HTF9C) knockout HeLa cell lysate

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TRMT2A 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 insertion in exon2.

View Alternative Names

HTF9C, HpaII tiny fragments locus 9c protein, TRM2 tRNA methyltransferase 2 homolog A, TRM2 tRNA methyltransferase 2 homolog A (S. cerevisiae), TRM2A_HUMAN, TRMT2A, tRNA (uracil-5-)-methyltransferase homolog A, tRNA methyltransferase 2 homolog A, tRNA methyltransferase 2, S. cerevisiae, homolog of, A

3 Images
Western blot - Human TRMT2A (HTF9C) knockout HeLa cell lysate (AB258253)
  • WB

Lab

Western blot - Human TRMT2A (HTF9C) knockout HeLa cell lysate (AB258253)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : TRMT2A knockout HeLa cell lysate (20 ug)
Lane 3 : MOLT-4 cell lysate (20 ug)

ab230922 was shown to specifically react with HTF9C/TRMT2A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265160 (knockout cell lysate ab258253) was used. Wild-type and HTF9C/TRMT2A knockout samples were subjected to SDS-PAGE. ab230922 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HTF9C/TRMT2A antibody [1G8] (<a href='/en-us/products/primary-antibodies/htf9c-trmt2a-antibody-1g8-ab230922'>ab230922</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TRMT2A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TRMT2A (HTF9C) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-trmt2a-htf9c-knockout-hela-cell-line-ab265160'>ab265160</a>)

Lane 3:

MOLT-4 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 69 kDa

Observed band size: 75 kDa

false

Western blot - Human TRMT2A (HTF9C) knockout HeLa cell lysate (AB258253)
  • WB

Lab

Western blot - Human TRMT2A (HTF9C) knockout HeLa cell lysate (AB258253)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : TRMT2A knockout HeLa cell lysate (20 ug)
Lane 3 : MOLT-4 cell lysate (20 ug)

ab205616 was shown to specifically react with HTF9C/TRMT2A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265160 (knockout cell lysate ab258253) was used. Wild-type and HTF9C/TRMT2A knockout samples were subjected to SDS-PAGE. ab205616 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-HTF9C/TRMT2A antibody [EPR16485-54] (<a href='/en-us/products/primary-antibodies/htf9c-trmt2a-antibody-epr16485-54-ab205616'>ab205616</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TRMT2A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TRMT2A (HTF9C) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-trmt2a-htf9c-knockout-hela-cell-line-ab265160'>ab265160</a>)

Lane 3:

MOLT-4 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 1/10000 dilution

Predicted band size: 69 kDa

Observed band size: 75 kDa

false

Sanger Sequencing - Human TRMT2A (HTF9C) knockout HeLa cell lysate (AB258253)
  • Sanger seq

Unknown

Sanger Sequencing - Human TRMT2A (HTF9C) knockout HeLa cell lysate (AB258253)

Homozygous : 1 bp insertion 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 insertion in exon2.

Disease

Adenocarcinoma

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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, 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?

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Properties and storage information

Gene name
TRMT2A
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.

HTF9C also known as TRMT2A is a tRNA methyltransferase enzyme with a mass of approximately 57 kDa. Its primary function is to catalyze the methylation of tRNA specifically the modification of uridine at position 54 to 5-methyluridine. This modification is essential for maintaining the correct structure and function of tRNA which affects protein synthesis. The protein is ubiquitously expressed in various tissues suggesting it plays an important role in fundamental cellular processes.
Biological function summary

The enzyme influences accurate tRNA function affecting overall protein translation efficiency. HTF9C/TRMT2A contributes to the maturation of tRNA molecules an important step in the stability and proper functioning of the translation machinery. Although it functions largely as an independent enzyme it may transiently interact with other proteins in the tRNA modification pathway indicating its potential involvement in broader RNA processing complexes.

Pathways

HTF9C/TRMT2A plays a critical role in the tRNA modification pathway which is tightly linked to protein biosynthesis. Its function affects the fidelity of translation connecting it indirectly to the mTOR pathway which regulates cell growth and metabolism. The TRMT2A enzyme's activity is potentially interrelated with ribosomal proteins like RPLP0 which are important for the translation process reflecting its impact on cellular growth and proliferation.

Improper function of HTF9C/TRMT2A can contribute to the development of cancer due to dysregulated protein synthesis. Aberrant methylation patterns affecting tRNA can result in translation anomalies contributing to oncogenic transformation. Additionally its malfunction is thought to be connected to neurological disorders where precise protein translation is important for neural function. In these contexts HTF9C may interact with dysregulated proteins such as p53 involved in cell cycle control and tumor suppression further illustrating its role in disease mechanisms.
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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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Complementary Products

Primary Antibodies

AB205616

Anti-HTF9C/TRMT2A antibody [EPR16485-54]

RabMAb|Recombinant|KO Validated

Western blot - Anti-HTF9C/TRMT2A antibody [EPR16485-54] (AB205616)

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Cell Lines & Lysates

AB265160

Human TRMT2A (HTF9C) knockout HeLa cell line

Advanced Validation

Western blot - Human TRMT2A (HTF9C) knockout HeLa cell line (AB265160)

  • 0
  • 0

Product promise

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