Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Neurofibromin antibody [EPR22989-68] - BSA and Azide free (ab260004)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22989-68] to Neurofibromin - BSA and Azide free
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Neurofibromin antibody [EPR22989-68] - BSA and Azide free
    See all Neurofibromin primary antibodies

  • Description

    Rabbit monoclonal [EPR22989-68] to Neurofibromin - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: Wild-type HAP1 and HeLa cell lysates.

  • General notes

    ab260004 is the carrier-free version of ab264070. 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

    ab260004 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.

Properties

Images

  • Western blot - Anti-Neurofibromin antibody [EPR22989-68] - BSA and Azide free (ab260004)
    Western blot - Anti-Neurofibromin antibody [EPR22989-68] - BSA and Azide free (ab260004)
    All lanes : Anti-Neurofibromin antibody [EPR22989-68] (ab238142) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : NFIB knockout HeLa cell lysate
    Lane 3 : HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 319 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab238142).

    Lanes 1-3: Merged signal (red and green). Green - ab238142 observed at 319 kDa. Red - loading control ab7291 observed at 50 kDa.

     ab238142 Recombinant Anti-NF1 antibody [EPR22989-68] was shown to specifically react with NFIB in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264725 (knockout cell lysate ab258533) was used. Wild-type and NF1 knockout samples were subjected to SDS-PAGE. ab238142 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution 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.

     

     

  • Western blot - Anti-Neurofibromin antibody [EPR22989-68] - BSA and Azide free (ab260004)
    Western blot - Anti-Neurofibromin antibody [EPR22989-68] - BSA and Azide free (ab260004)
    All lanes : Anti-Neurofibromin antibody [EPR22989-68] (ab238142) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : Neurofibromin knockout HAP1 whole cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 319 kDa
    Observed band size: 26 kDa
    why is the actual band size different from the predicted?



    ab238142 was shown to specifically react with Neurofibromin in wild-type HAP1 cells as signal was lost in Neurofibromin knockout cells. Wild-type and Neurofibromin knockout samples were subjected to SDS-PAGE. ab238142 and ab129002 (Rabbit anti-Vinculin loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/5000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique. Lysate should be made freshly and used in WB immediately to minimize protein degradation. Degraded fragment(250 KD) observed is consistent with what has been described in the literature (PMID: 30131853, PMID: 30408279).

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

    Exposure time: 26 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab238142).

  • Anti-Neurofibromin antibody [EPR22989-68] - BSA and Azide free (ab260004)
    Anti-Neurofibromin antibody [EPR22989-68] - BSA and Azide free (ab260004)

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