Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)

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 - BSA and Azide free
  • Suitable for: IP, WB, Flow Cyt, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Rat, Human

Overview

  • Product name

    Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free
    See all NDUFS3 primary antibodies

  • Description

    Rabbit monoclonal [EPR12782] to NDUFS3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • HEK293T, HepG2, 293T, HL60 and fetal heart lysates. Human heart and kidney tissue. 293 cells.

  • General notes

    Ab249976 is the carrier-free version of ab177471. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab249976 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Images

  • Western blot - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    Western blot - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    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?



    This data was developed using the same antibody clone in a different buffer formulation (ab177471).

    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 - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    Immunocytochemistry - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)

    This data was developed using ab177471, the same antibody clone in a different buffer formulation.

    Immunofluorescence analysis of 293 cells labeling NDUFS3 with ab177471 at 1/50 dilution (green). DAPI nuclear staining (blue).
  • Flow Cytometry - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    Flow Cytometry - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    This data was developed using ab177471, the same antibody clone in a different buffer formulation.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).
  • Immunoprecipitation - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    Immunoprecipitation - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)

    This data was developed using ab177471&

    44; the same antibody clone in a different buffer formulation.

    >Western blot analysis on immunoprecipitation pellet from (1) human fetal heart lysate or (2) 1XPBS (negative control) using ab177471 at 1/10 dilution, and HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.
  • Western blot - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    Western blot - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    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



    This data was developed using the same antibody clone in a different buffer formulation (ab177471).

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    This data was developed using ab177471, the same antibody clone in a different buffer formulation.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] - BSA and Azide free (ab249976)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    This data was developed using ab177471, the same antibody clone in a different buffer formulation.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.
  • Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
  • Western blot - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    Western blot - Anti-NDUFS3 antibody [EPR12782] - BSA and Azide free (ab249976)
    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



    This data was developed using ab177471, the same antibody clone in a different buffer formulation.

     

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