Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17939] to HADHA - BSA and Azide free
  • Suitable for: ICC/IF, IP, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-HADHA antibody [EPR17939] - BSA and Azide free
    See all HADHA primary antibodies

  • Description

    Rabbit monoclonal [EPR17939] to HADHA - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IP, Flow Cyt, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HeLa, Jurkat, HEK293, SH-SY5Y and HepG2 whole cell lysates; Human fetal brain, fetal kidney and fetal liver lysates; Mouse kidney, rat heart and rat kidney lysates. ICC/IF: Jurkat and HeLa cells. IP: HEK293 whole cell lysate. Flow Cyt: Jurkat (human acute T cell leukemia).

  • General notes

    Ab242411 is the carrier-free version of ab200652. 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

    ab242411 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.

Images

  • Western blot - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)
    Western blot - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)
    All lanes : Anti-HADHA antibody [EPR17939] (ab200652) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : H2AFY Knockout HEK-293T cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : SH-SY5Y cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 83 kDa
    Observed band size: 82 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 (ab200652).

    Lanes 1-4: Merged signal (red and green). Green - ab200652 observed at 82 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab200652 Anti-HADHA antibody [EPR17939] was shown to specifically react with HADHA in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266274 (knockout cell lysate ab257464) was used. Wild-type and HADHA knockout samples were subjected to SDS-PAGE. ab200652 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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.

     

     

  • Flow Cytometry - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)
    Flow Cytometry - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)

    ab200652 staining HADHA in Jurkat (human acute T cell leukemia) cells by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/2200. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200652)
  • Immunocytochemistry/ Immunofluorescence - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)
    Immunocytochemistry/ Immunofluorescence - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling HADHA with ab200652 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Cytoplasm staining on Jurkat cell line is observed.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab200652 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200652)

  • Western blot - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)
    Western blot - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)
    All lanes : Anti-HADHA antibody [EPR17939] (ab200652) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : HADHA knockout HAP1 cell lysate
    Lane 3 : HEK293 cell lysate
    Lane 4 : HepG2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 83 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab200652 observed at 82 kDa. Red - loading control, ab8245, observed at 37 kDa.


    ab200652 was shown to specifically react with HADHA when HADHA knockout samples were used. Wild-type and HADHA knockout samples were subjected to SDS-PAGE. ab200652 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200652)

  • Immunocytochemistry/ Immunofluorescence - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)
    Immunocytochemistry/ Immunofluorescence - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling HADHA with ab200652 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Cytoplasm staining on HeLa cell line is observed.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab200652 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200652)

  • Immunoprecipitation - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)
    Immunoprecipitation - Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)

    HADHA was immunoprecipitated from 1mg of HEK293 (Human embryonic kidney) whole cell lysate with ab200652 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab200652 at 1/2000 dilution.

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: HEK293 whole cell lysate 10 µg (Input).

    Lane 2: ab200652 IP in HEK293 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200652 in HEK293 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200652)
  • Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)
    Anti-HADHA antibody [EPR17939] - BSA and Azide free (ab242411)

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