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AB266419

Human NDUFS3 knockout HEK-293T cell line

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NDUFS3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

CI 30, CI-30kD, COMPLEX I, MITOCHONDRIAL RESPIRATORY CHAIN, 30-KD SUBUNIT, Complex I 30kDa subunit, Complex I-30kD, NADH coenzyme Q reductase, NADH dehydrogenase (ubiquinone) Fe S protein 3 30kDa, NADH dehydrogenase [ubiquinone] iron sulfur protein 3 mitochondrial, NADH dehydrogenase [ubiquinone] iron-sulfur protein 3, NADH dehydrogenase ubiquinone 30 kDa subunit, NADH-Ubiquinone Oxidoreductase Fe-S Protein 3, NADH-ubiquinone oxidoreductase 30 kDa subunit, NDUS3_HUMAN, mitochondrial

5 Images
Western blot - Human NDUFS3 knockout HEK-293T cell line (AB266419)
  • WB

Lab

Western blot - Human NDUFS3 knockout HEK-293T cell line (AB266419)

Lanes 1 - 2 : Merged signal (red and green). Green - ab177471 observed at 27 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab177471 was shown to react with NDUFS3 in HEK-293T wild-type cells in western blot with loss of signal observed in NDUFS3 knockout cell line ab266419 (NDUFS3 knockout cell lysate ab257556). Wild-type and NDUFS3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab177471 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (<a href='/en-us/products/primary-antibodies/ndufs3-antibody-epr12782-c-terminal-ab177471'>ab177471</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

NDUFS3 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human NDUFS3 knockout HEK-293T cell line (ab266419)

Predicted band size: 30 kDa

Observed band size: 27 kDa

false

Western blot - Human NDUFS3 knockout HEK-293T cell line (AB266419)
  • WB

Lab

Western blot - Human NDUFS3 knockout HEK-293T cell line (AB266419)

Lanes 1- 4 : Merged signal (red and green). Green - ab183733 observed at 30 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab183733 was shown to react with NDUFS3 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266419 (knockout cell lysate ab257556) was used. Wild-type HEK-293T and NDUFS3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab183733 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4° at a 1 in 10000 dilution and a 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-NDUFS3 antibody [EPR12781] (<a href='/en-us/products/primary-antibodies/ndufs3-antibody-epr12781-ab183733'>ab183733</a>) at 1/10000 dilution

Lane 1:

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

Lane 2:

NDUFS3 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human NDUFS3 knockout HEK-293T cell line (ab266419)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HL60 cell lysate at 20 µg

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

Western blot - Human NDUFS3 knockout HEK-293T cell line (AB266419)
  • WB

Lab

Western blot - Human NDUFS3 knockout HEK-293T cell line (AB266419)

Lanes 1 - 2 : Merged signal (red and green). Green - ab183733 observed at 27 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab183733 was shown to react with NDUFS3 in HEK-293T wild-type cells in western blot with loss of signal observed in NDUFS3 knockout cell line ab266419 (NDUFS3 knockout cell lysate ab257556). Wild-type and NDUFS3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab183733 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4° at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NDUFS3 antibody [EPR12781] (<a href='/en-us/products/primary-antibodies/ndufs3-antibody-epr12781-ab183733'>ab183733</a>) at 1/10000 dilution

Lane 1:

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

Lane 2:

NDUFS3 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human NDUFS3 knockout HEK-293T cell line (ab266419)

Predicted band size: 30 kDa

Observed band size: 27 kDa

false

Western blot - Human NDUFS3 knockout HEK-293T cell line (AB266419)
  • WB

Lab

Western blot - Human NDUFS3 knockout HEK-293T cell line (AB266419)

Lanes 1- 4 : Merged signal (red and green). Green - ab177471 observed at 30 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab177471 was shown to react with NDUFS3 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266419 (knockout cell lysate ab257556) was used. Wild-type HEK-293T and NDUFS3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab177471 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4° at a 1 in 1000 dilution and a 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-NDUFS3 antibody [EPR12782] - C-terminal (<a href='/en-us/products/primary-antibodies/ndufs3-antibody-epr12782-c-terminal-ab177471'>ab177471</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

NDUFS3 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human NDUFS3 knockout HEK-293T cell line (ab266419)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HL60 cell lysate at 20 µg

Predicted band size: 30 kDa

Observed band size: 30 kDa

false

Sanger Sequencing - Human NDUFS3 knockout HEK-293T cell line (AB266419)
  • Sanger seq

Unknown

Sanger Sequencing - Human NDUFS3 knockout HEK-293T cell line (AB266419)

Homozygous : 19 bp deletion in exon 1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 1

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Reactivity data

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

We will provide viable cells that proliferate on revival.

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.

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What's included?

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Properties and storage information

Gene name
NDUFS3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

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.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

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

NADH:ubiquinone oxidoreductase core subunit S3 (NDUFS3) is an important component of the mitochondrial respiratory chain complex I also known as NADH:ubiquinone oxidoreductase. With an approximate mass of 25 kDa NDUFS3 plays an integral role in the assembly and function of complex I. This protein is mostly expressed in the mitochondria across various tissues. It is better known as a core subunit important for the catalytic activity of complex I.
Biological function summary

NDUFS3 participates in cellular respiration by facilitating the electron transfer from NADH to ubiquinone. It is part of the complex I assembly that comprises 45 different subunits. This large assembly is the first enzyme of the mitochondrial electron transport chain and it ensures efficient energy production in the form of ATP. In its role NDUFS3 collaborates closely with other core subunits like NDUFS1 and NDUFS2 to maintain the proper function of cellular metabolism.

Pathways

NDUFS3 is essential in the oxidative phosphorylation pathway which plays an important part in ATP generation. By interacting with other complex I subunits NDUFS3 enables electron flow that drives ATP synthase activity. Additionally it is connected to the apoptosis pathway. Its dysfunction may result in disrupted energy metabolism which can trigger cell death. NDUFS3 also interacts with proteins like NDUFV1 and NDUFV2 highlighting its centrality in energy and signal transduction processes.

NDUFS3 links to mitochondrial disorders and neurodegenerative diseases such as Leigh syndrome. Mutations in NDUFS3 can impair mitochondrial function leading to reduced ATP production and accumulation of defective mitochondria. These alterations contribute to the progressive deterioration seen in these disorders. In the context of neurodegenerative diseases NDUFS3 dysfunction may associate with proteins like cytochrome c emphasizing its role in mitochondria-dependent apoptotic pathways and neuronal 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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Target data

See full target information NDUFS3
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Product promise

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