Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4686-66] to NFkB p100/NFKB2 - BSA and Azide free
  • Suitable for: ICC, WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free
    See all NFkB p100/NFKB2 primary antibodies

  • Description

    Rabbit monoclonal [EPR4686-66] to NFkB p100/NFKB2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, WB, IPmore details
    Unsuitable for: Flow Cyt or IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human NFkB p100/NFKB2. The exact sequence is proprietary.
    Database link: Q00653

  • Positive control

    • WB: Jurkat, HeLa, Daudi, HCT116 and MCF7 cell lysates. ICC/IF: Wild-type HAP1 cells.

  • General notes

    ab249847 is the carrier-free version of ab175192 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

    Ab249847 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product was previously labelled as NFkB p100

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)

    This data was developed using ab175192, the same antibody clone in a different buffer formulation.

    Lane 1: Wild-type HAP1 cell lysate (20 µg)

    Lane 2: NFkB p100 knockout HAP1 cell lysate (20 µg)

    Lane 3: Jurkat cell lysate (20 µg)

    Lane 4: HeLa cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab175192 observed at 97 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab175192 was shown to specifically react with NF?B p100 when NF?B p100 knockout samples were used. Wild-type and NF?B p100 knockout samples were subjected to SDS-PAGE. ab175192 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 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 1/10000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry - Anti-NFkB p100 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    Immunocytochemistry - Anti-NFkB p100 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    This data was developed using ab175192, the same antibody clone in a different buffer formulation.ab175192 staining NFkB p100/NFKB2 in wild-type HAP1 cells (top panel) and NFkB p100/NFKB2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab175192 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
  • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    All lanes : Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192) at 1/1000 dilution

    Lane 1 : Wild-type HCT116 cell lysate
    Lane 2 : NFKB2 knockout HCT116 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 97 kDa
    Observed band size: 120 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 (ab175192).

    Lanes 1- 2: Merged signal (red and green). Green - ab175192 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab175192 was shown to react with NFkB p100/NFKB2 in wild-type HCT116 cells in western blot. The band observed in knockout cell line ab266883 (knockout cell lysate ab257245) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HCT116 and NFKB2 knockout HCT117 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. ab175192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 10000 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.

  • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    All lanes : Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192) at 1/1000 dilution

    Lane 1 : Wild-type HepG2 cell lysate
    Lane 2 : NFKB2 knockout HepG2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 97 kDa
    Observed band size: 120 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 containing PBS, BSA, glycerol, and sodium azide (ab175192).

    Lanes 1- 2: Merged signal (red and green). Green - ab175192 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab175192 was shown to react with NFkB p100/NFKB2 in wild-type HepG2 cells in western blot. The band observed in knockout cell line ab262323 (knockout cell lysate ab257247) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HepG2 and NFKB2 knockout HepG2 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. ab175192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 10000 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.

  • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    All lanes : Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192) at 1/10000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : Daudi cell lysates
    Lane 4 : MCF7 cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 97 kDa



    This data was developed using ab175192, the same antibody clone in a different buffer formulation.

  • Immunoprecipitation - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    Immunoprecipitation - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    This data was developed using ab175192, the same antibody clone in a different buffer formulation.Western blot analysis on immunoprecipitation pellet from Jurkat cell lysate labeling NFkB p100/NFKB2 with ab175192 at 1/10 dilution.
  • Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
    Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)

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