NDUFA13 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and 73 bp insertion in exon 2 and 8 bp deletion in exon 2.
2700054G14Rik, AU022060, B16.6, CDA016, CGI-39, CGI39 protein, CI-B16.6, Cell death regulatory protein, Cell death regulatory protein GRIM-19, Complex I-B16.6, FLJ58045, FLJ59191, Gene associated with retinoic and IFN-induced mortality 19 protein, Gene associated with retinoic and interferon-induced mortality 19 protein, Gene associated with retinoic interferon induced mortality 19 protein, NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, 13, NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13, NADH-ubiquinone oxidoreductase B16.6 subunit, NDUAD_HUMAN, NDUFA13, RGD1565358
NDUFA13 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 2 and 73 bp insertion in exon 2 and 8 bp deletion in exon 2.
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.
GRIM19 also known as NDUFA13 is a small protein with a molecular mass of approximately 16 kDa. It expresses mainly in mitochondria where it forms an integral component of the mitochondrial respiratory chain complex I. GRIM19 plays an important role in the functioning of complex I by facilitating the assembly and stabilization of the complex enabling efficient electron transport and ATP synthesis.
GRIM19 interacts with several complexes and proteins within the mitochondrial inner membrane. As a part of complex I also known as NADH:ubiquinone oxidoreductase it contributes to the initial step of mitochondrial oxidative phosphorylation. GRIM19 also participates in the regulation of apoptosis and its expression is influenced during cellular stress situations highlighting its significance in cellular survival and energy production.
GRIM19 integrates closely with mitochondrial apoptosis and energy metabolism pathways. Its interactions within oxidative phosphorylation place it in concert with both the electron transport chain and the apoptosis regulatory network. GRIM19 shares associations with related proteins such as cytochrome c oxidase and other complex I subunits highlighting its essential contribution to these important biological processes.
GRIM19 demonstrates links to various pathologies including cancer and mitochondrial diseases. Alterations in GRIM19 expression or function have been implicated in the progression of certain cancers such as colorectal and thyroid cancer where GRIM19's usual regulatory role in apoptosis can be disrupted. Additionally its involvement in mitochondrial disease connects to proteins like COX-A1 emphasizing the importance of maintaining functional complexes for cellular health.
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Lanes 1-3: Merged signal (red and green). Green - Anti-GRIM19 antibody [6E1BH7] ab110240 observed at 17 kDa. Red - loading control Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 observed at 36 kDa.
Anti-GRIM19 antibody [6E1BH7] ab110240 GRIM19 antibody [6E1BH7] was shown to specifically react with GRIM19 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265863 (knockout cell lysate Human NDUFA13 (GRIM19) knockout HeLa cell lysate ab257136) was used. Wild-type and GRIM19 knockout samples were subjected to SDS-PAGE. Anti-GRIM19 antibody [6E1BH7] ab110240 and Anti-GAPDH antibody[EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) 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® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (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-GRIM19 antibody [6E1BH7] (Anti-GRIM19 antibody [6E1BH7] ab110240) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: NDUFA13 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human NDUFA13 (GRIM19) knockout HeLa cell line (ab265863)
Lane 3: Jurkat cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution
Predicted band size: 17 kDa, 184 kDa, 28 kDa, 56 kDa, 62 kDa, 76 kDa
Observed band size: 17 kDa, 184 kDa, 29 kDa, 56 kDa, 70 kDa, 75 kDa
Lanes 1-3: Merged signal (red and green). Green - Anti-GRIM19 antibody [EPR4471(2)] ab109017 observed at 17 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-GRIM19 antibody [EPR4471(2)] ab109017 Anti-GRIM19 antibody [EPR4471(2)] was shown to specifically react with GRIM19 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265863 (knockout cell lysate Human NDUFA13 (GRIM19) knockout HeLa cell lysate ab257136) was used. Wild-type and GRIM19 knockout samples were subjected to SDS-PAGE. Anti-GRIM19 antibody [EPR4471(2)] ab109017 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-GRIM19 antibody [EPR4471(2)] (Anti-GRIM19 antibody [EPR4471(2)] ab109017) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: NDUFA13 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human NDUFA13 (GRIM19) knockout HeLa cell line (ab265863)
Lane 3: Jurkat 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: 17 kDa
Observed band size: 17 kDa
Allele-2: 2 bp deletion in exon 2.
Allele-3: 73 bp insertion in exon 2.
Allele-1: 8 bp deletion in exon 2.
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