WDR4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 5 and 94 bp insertion in exon 5.
OTTHUMP00000109401, OTTHUMP00000109402, TRM82, WD repeat protein 4, WD repeat protein domain 4 protein isoform 2, WD repeat-containing protein 4, WDR4_HUMAN, tRNA (guanine-N(7)-)-methyltransferase subunit WDR4
WDR4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 5 and 94 bp insertion in exon 5.
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.
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.
The WDR4 protein also known as WD repeat domain 4 functions in the modification of ribonucleic acids. It specifically takes part in the methylation process of N(7)-methylguanine-tRNA (m7G46) where it serves as an important component of the tRNA methyltransferase complex. With an approximate mass of 51 kDa WDR4 shows expression in various tissues throughout the human body contributing to fundamental cellular processes.
WDR4 plays a role in the maintenance of accurate protein synthesis. It contributes to the stabilization and correct folding of tRNA molecules which are essential for the translation of genetic information from mRNA to proteins. As part of the tRNA methyltransferase complex WDR4 interacts with the METTL1 enzyme highlighting its involvement in precise tRNA modification processes critical for cellular function and proliferation.
The function of WDR4 integrates into the broader landscape of the RNA modification pathway. This pathway is tied to the regulation of gene expression and cellular homeostasis. WDR4's activity impacts the efficiency and fidelity of the translation pathway. In this context WDR4 works closely with METTL1 enhancing the stability and functional capacity of tRNA during protein synthesis which is pivotal for the correct transmission of genetic code within cells.
WDR4's malfunction or altered expression associates with several conditions including microcephalic primordial dwarfism where its disruption affects brain development and growth. The protein interacts with METTL1 implicated in the same disorder demonstrating the importance of their coordinated function. Additionally alterations in WDR4 expression might link to certain types of cancers where aberrant tRNA methylation affects tumor growth and progression indicating the potential for WDR4 as a therapeutic target.
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Lanes 1-3: Merged signal (red and green). Green - Anti-WDR4 antibody [EPR11052] ab169526 observed at 49 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-WDR4 antibody [EPR11052] ab169526 Anti-WDR4 antibody [EPR11052] was shown to specifically react with WDR4 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265109 (knockout cell lysate Human WDR4 knockout HeLa cell lysate ab258285) was used. Wild-type and WDR4 knockout samples were subjected to SDS-PAGE. Anti-WDR4 antibody [EPR11052] ab169526 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-WDR4 antibody [EPR11052] (Anti-WDR4 antibody [EPR11052] ab169526) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: WDR4 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human WDR4 knockout HeLa cell line (ab265109)
Lane 3: HepG2 cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 45 kDa
Observed band size: 49 kDa
Allele-2: 94 bp insertion in exon 5.
Allele-1: 1 bp insertion in exon 5.
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