Antibodies, Proteins, Kits and Reagents for Life Science

314 937 ₸ Product size

100 µl 314 937 ₸

Order now and get it on Tuesday March 02, 2021

Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR12782] to NDUFS3 - C-terminal
Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, IP
Knockout validated
Reacts with: Human

Product name

Anti-NDUFS3 antibody [EPR12782] - C-terminal
See all NDUFS3 primary antibodies
Description

Rabbit monoclonal [EPR12782] to NDUFS3 - C-terminal
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human NDUFS3 aa 200 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
Database link: O75489
Positive control
- HepG2, 293T, HL60 and fetal heart lysates. Human heart and kidney tissue. 293 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR12782
Isotype

IgG
Research areas

Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

All lanes : Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : NDUFS3 knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 30 kDa
Observed band size: 27 kDa
why is the actual band size different from the predicted?

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°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.
Immunocytochemistry/ Immunofluorescence - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

Immunofluorescence analysis of 293 cells labeling NDUFS3 with ab177471 at 1/50 dilution (green). DAPI nuclear staining (blue).
Flow Cytometry - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

Flow cytometry analysis of HEK293 (Human embryonic kidney epithelial cell) cells labeling NDUFS3 (red) with purified ab177471 at a 1/200 dilution (10ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluor^®488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).
Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471) Image is courtesy of an anonymous Abreview.

All lanes : Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471) at 1/1000 dilution

Lanes 1-2 : Whole cell lysates from NDUFS3 KO cells.
Lanes 3-4 : Whole cell lysates from NDUFS3 WT cells.
Lanes 5-6 : Isolated mitochondria from NDUFS3 KO Cells.
Lanes 7-8 : Isolated mitochondria from NDUFS3 WT Cells.

Lysates/proteins at 50 µg per lane.

Secondary
All lanes : Goat anti-Rabbit polyclonal HRP conjugate. at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 30 kDa

Exposure time: 1 minute
See Abreview
Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

All lanes : Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : NDUFS3 knockout HEK-293T cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HL60 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 30 kDa
Observed band size: 30 kDa

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°C 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.
Immunoprecipitation - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

Western blot analysis on immunoprecipitation pellet from (1) human fetal heart lysate or (2) 1XPBS (negative control) using ab177471at 1/10 dilution, and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

Immunohistochemcal analysis of paraffin-embedded Human heart tissue labeling NDUFS3 with ab177471 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

Immunohistochemcal analysis of paraffin-embedded Human kidney tissue labeling NDUFS3 with ab177471 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Western blot - Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

All lanes : Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471) at 1/1000 dilution

Lane 1 : HepG2 lysate
Lane 2 : 293T lysate
Lane 3 : HL60 lysate
Lane 4 : Fetal heart lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat-anti-rabbit HRP at 1/2000 dilution

Developed using the ECL technique.

Predicted band size: 30 kDa
Anti-NDUFS3 antibody [EPR12782] - C-terminal (ab177471)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com