JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB257556

Human NDUFS3 knockout HEK-293T cell lysate

Be the first to review this product! Submit a review

|

(0 Publication)

NDUFS3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 1.

View Alternative Names

Complex I-30kD, NDUFS3, CI-30kD, NADH-ubiquinone oxidoreductase 30 kDa subunit

5 Images
Western blot - Human NDUFS3 knockout HEK-293T cell lysate (AB257556)
  • WB

Lab

Western blot - Human NDUFS3 knockout HEK-293T cell lysate (AB257556)

Lane 1 : Wild-type HEK-293T cell lysate 20 ug
Lane 2 : NDUFS3 knockout HEK-293T cell lysate 20 ug
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 wild-type HEK-293T 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°C 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 (<a href='/en-us/products/cell-lines/human-ndufs3-knockout-hek-293t-cell-line-ab266419'>ab266419</a>)

Predicted band size: 30 kDa

Observed band size: 27 kDa

false

Western blot - Human NDUFS3 knockout HEK-293T cell lysate (AB257556)
  • WB

Lab

Western blot - Human NDUFS3 knockout HEK-293T cell lysate (AB257556)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : NDUFS3 knockout HEK-293T cell lysate (20µg)

Lane 3 : HepG2 cell lysate (20µg)

Lane 4 : HL60 cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab183733 observed at 30 kDa. Red - loading control ab7291 observed at 50 kDa.

ab183733 Rabbit monoclonal [EPR12781] to NDUFS3 was shown to specifically 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 and NDUFS3 knockout samples 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) were incubated overnight at 4°C at 1 in 10000 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-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 (<a href='/en-us/products/cell-lines/human-ndufs3-knockout-hek-293t-cell-line-ab266419'>ab266419</a>)

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 lysate (AB257556)
  • WB

Lab

Western blot - Human NDUFS3 knockout HEK-293T cell lysate (AB257556)

Lane 1 : Wild-type HEK-293T cell lysate 20 ug
Lane 2 : NDUFS3 knockout HEK-293T cell lysate 20 ug
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 wild-type HEK-293T 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°C 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 (<a href='/en-us/products/cell-lines/human-ndufs3-knockout-hek-293t-cell-line-ab266419'>ab266419</a>)

Predicted band size: 30 kDa

Observed band size: 27 kDa

false

Western blot - Human NDUFS3 knockout HEK-293T cell lysate (AB257556)
  • WB

Lab

Western blot - Human NDUFS3 knockout HEK-293T cell lysate (AB257556)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : NDUFS3 knockout HEK-293T cell lysate (20µg)

Lane 3 : HepG2 cell lysate (20µg)

Lane 4 : HL60 cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab177471 observed at 30 kDa. Red - loading control ab7291 observed at 50 kDa.

ab177471 Rabbit monoclonal [EPR12782] to NDUFS3 - C-terminal was shown to specifically 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 and NDUFS3 knockout samples 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) 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 (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 (<a href='/en-us/products/cell-lines/human-ndufs3-knockout-hek-293t-cell-line-ab266419'>ab266419</a>)

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 lysate (AB257556)
  • Sanger seq

Unknown

Sanger Sequencing - Human NDUFS3 knockout HEK-293T cell lysate (AB257556)

Homozygous : 19 bp deletion in exon 1

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, Homozygous: 19 bp deletion in exon 1.

style
left-column

Complementary Products

Primary Antibodies

AB14711

Anti-NDUFS3 antibody [17D95]

Flow Cytometry - Anti-NDUFS3 antibody [17D95] (AB14711)

  • 4
  • 1
Cell Lines & Lysates

AB266419

Human NDUFS3 knockout HEK-293T cell line

Advanced Validation

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

  • 0
  • 0
Proteins & Peptides

AB136706

Recombinant Human NDUFS3 protein (His tag N-Terminus)

SDS-PAGE - Recombinant Human NDUFS3 protein (His tag N-Terminus) (AB136706)

  • 0
  • 0
style
left-column

Reactivity data

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

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 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.

style
left-column

What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257556-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human NDUFS3 knockout HEK293T cell lysate", "number":"AB257556-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB257556-CMP02", "productcode":"" } ] } } }
style
left-column

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
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
style
left-column

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.
style
left-column

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

style
left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

style
left-column

Product protocols

style
left-column
style
left-column
style
right-column

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'.

For licensing inquiries, please contact partnerships@abcam.com