Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1238Y] to Notch1 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Notch1 antibody [EP1238Y] - BSA and Azide free
    See all Notch1 primary antibodies

  • Description

    Rabbit monoclonal [EP1238Y] to Notch1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Cow

  • Immunogen

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

  • Positive control

    • WB: HeLa, HEK293, HAP1 and MOLT-4 cell lysate. IHC-P: Human brain tissue. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.

  • General notes

    Ab221603 is the carrier-free version of ab52627. 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

    ab221603 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

Images

  • Western blot - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
    Western blot - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
    All lanes : Anti-Notch1 antibody [EP1238Y] (ab52627) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : NOTCH1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Observed band size: 110 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 (ab52627).

    Lanes 1- 2: Merged signal (red and green). Green - ab52627 observed at 110 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab52627 was shown to react with Notch1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261762 (knockout cell lysate ab257006) was used. Wild-type HeLa and NOTCH1 knockout HeLa 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. ab52627 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)

    Immunohistochemical staining of paraffin-embedded human brain with purified ab52627 at a dilution of 1/150. A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627).

  • Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
    Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)

    Immunofluorescent staining of HeLa cells fixed with 4% PFA using purified ab52627 at a dilution of 1/150. An Alexa Fluor® 555 goat anti-rabbit was used as the secondary and the sample was stained with DAPI. An Alexa Fluor® 555 goat anti-mouse was used at a dilution of 1/500 after ab52627 as the negative control and is shown in the bottom right hand panel.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627).

  • Flow Cytometry - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
    Flow Cytometry - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)

    Flow cytometry analysis of permeabilized HeLa cells (2% PFA, pink) with purified ab52627 at a 1/200 dilution, or negative control rabbit monoclonal IgG (green). The secondary antibody was FITC goat anti-rabbit.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627).

  • Western blot - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
    Western blot - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
    All lanes : Anti-Notch1 antibody [EP1238Y] (ab52627) at 1 µg/ml

    Lane 1 : Wild-type HAP1 cell lysate at 40 µg
    Lane 2 : NOTCH1 knockout HAP1 cell lysate at 40 µg
    Lane 3 : MOLT-4 cell lysate at 20 µg
    Lane 4 : HepG2 cell lysate at 20 µg

    Performed under reducing conditions.

    Observed band size: 105 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 (ab52627).

    Lanes 1 - 4: Merged signal (red and green). Green - ab52627 observed at 105 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab52627 was shown to react with Notch1 in wild-type HAP1 cells in western blot. Loss of signal was observed when NOTCH1 knockout sample was used. Wild-type and NOTCH1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with ab52627 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 µg/ml 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)

    Immunohistochemical staining of paraffin-embedded human brain with unpurified ab52627 at a dilution of 1/100. A prediluted HRP polymer for rabbit IgG was used as the secondary and the sample was stained with hematoxylin. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627).

  • Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
    Immunocytochemistry/ Immunofluorescence - Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)

    Immunofluorescent staining of HeLa cells fixed with 4% PFA using unpurified ab52627 at a dilution of 1/100. An Alexa Fluor® 555 goat anti-rabbit was used as the secondary and the sample was stained with DAPI. An Alexa Fluor® 555 goat anti-mouse was used at a dilution of 1/500 as the negative control and is shown in the bottom right hand panel.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52627).

  • Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)
    Anti-Notch1 antibody [EP1238Y] - BSA and Azide free (ab221603)

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