Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16629] to IKK gamma/NEMO - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free
    See all IKK gamma/NEMO primary antibodies

  • Description

    Rabbit monoclonal [EPR16629] to IKK gamma/NEMO - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Mouse
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal brain, Human fetal kidney, Human colon cancer, HEK293 whole cell lysates, HeLa whole cell lysates, K562 whole cell lysates, Jurkat whole cell lysates. Mouse brain, heart, kidney and spleen. Rat brain, heart, kidney and spleen. C6 whole cell lysates, RAW 264.7 whole cell lysates, PC-12 whole cell lysates, NIH/3T3 whole cell lysates. IHC-P: Human colonic adenocarcinoma, rat colon. ICC/IF: HeLa, NIH/3T3

  • General notes

    Ab230832 is the carrier-free version of ab178872. 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

    ab230832 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

Images

  • Western blot - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)
    Western blot - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)

    This WB data was generated using the same anti-IKK gamma antibody clone, EPR16629, in a different buffer formulation (cat# ab178872).

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: IKK gamma/NEMO knockout HAP1 cell lysate (20 µg) 
    Lane 3: Jurkat cell lysate (20 µg)
    Lane 4: K562 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab178872 observed at 40, 45, 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab178872 was shown to react with IKK gamma in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when IKK gamma/NEMO knockout samples were examined. Wild-type and IKK gamma/NEMO knockout samples were subjected to SDS-PAGE. ab178872 and ab8245 (loading control to GAPDH) were diluted at 1/5000 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 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)
    Immunocytochemistry/ Immunofluorescence - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)

    Immunofluorescence analysis of 4% paraformaldehyde fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells (Mouse embyro fibroblast cells) labeling IKK gamma/NEMO (green) with ab178872 at 1/250 dilution showing cytoplasm and nucleus staining. Secondary ab: Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/200 dilution. Counter stain is labeling tubulin (red) with ab7291 at 1/500 dilution with secondary antibody Goat anti-Mouse AlexaFluor® 594 (ab150120) at 1/400 dilution. DAPI stains the nucleus in blue. -ve control 1 is ab178872 at 1/250 dilution, ab150120 at 1/400 dilution. -ve control 2 is ab7291 at 1/500 dilution, ab150077 at 1/200 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178872).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)

    This IHC data was generated using the same anti-IKK gamma/NEMO antibody clone, EPR16629, in a different buffer formulation (cat# ab178872).

    Immunohistochemical analysis of paraffin embedded human colonic adenocarcinoma tissue labeling IKK gamma with ab178872 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic staining on colonic adenocarcinoma is observed.

    Negative control: Using PBS instead of primary ab, secondary ab ImmunoHistoprobe (Ready to use) HRP Polymer for Rabbit/Mouse IgG.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)
    Immunoprecipitation - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)

    IKK gamma/NEMO was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab178872 at 1/50 dilution. Western blot was performed of the immunoprecipitate using ab178872 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Left lane: HeLa whole cell extract. Right lane:  PBS instead of HeLa whole cell extract.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting 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 (ab178872).

  • Western blot - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)
    Western blot - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)
    All lanes : Anti-IKK gamma/NEMO antibody [EPR16629] (ab178872) at 1/1000 dilution

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

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 48 kDa
    Observed band size: 48 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab178872).

      Lanes 1- 2: Merged signal (red and green). Green - ab178872 observed at 48 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab178872 was shown to react with IKK gamma/NEMO in wild-type HEK-293T cells in western blot. The band observed in knockout cell line ab266674 (knockout cell lysate ab257153) lane below 48kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HEK-293T and IKBKG 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. ab178872 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)
    Immunocytochemistry/ Immunofluorescence - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)

    This ICC data was generated using the same anti-IKK gamma/NEMO antibody clone, EPR16629, in a different buffer formulation (cat# ab178872).

    Immunofluorescence analysis of 4% paraformaldehyde fixed, 0.1% Triton X-100 permeabilized HeLa cells (Human epithelial cells from cervix adenocarcinoma) labeling IKK gamma (green) with ab178872 at 1/250 dilution showing cytoplasm and nucleus staining. Secondary ab: Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/200 dilution. Counter stain is labeling tubulin (red) with ab7291 at 1/500 dilution with secondary antibody Goat anti-Mouse AlexaFluor® 594 (ab150120) at 1/400 dilution. DAPI stains the nucleus in blue. -ve control 1 is ab178872 at 1/250 dilution, ab150120 at 1/400 dilution. -ve control 2 is ab7291 at 1/500 dilution, ab150077 at 1/200 dilution.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)

    Immunohistochemical analysis of paraffin embedded Rat colon tissue labeling IKK gamma/NEMO with ab178872 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Counter stain is Hematoxylin. Cytoplasm staining on epithelial cells of rat colon is observed.

    Negative control: Using PBS instead of primary ab, secondary ab as above.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178872).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)
    Anti-IKK gamma/NEMO antibody [EPR16629] - BSA and Azide free (ab230832)

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