Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5777] to HP1 alpha - BSA and Azide free
  • Suitable for: WB, IP, IHC-P, Flow Cyt, ICC
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free
    See all HP1 alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR5777] to HP1 alpha - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC
    Human
    IHC-P
    Rat
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • ICC: HAP1 and HeLa cells; IP: MCF7 cell lysate; Flow Cyt: HeLa cells; IHC-P: Human breast cancer, rat pancreas, and mouse kidney tissue sections; WB: HeLa, SH-SY5Y whole cell lysates, Rat and mouse brain lysates.

  • General notes

    Ab226049 is the carrier-free version of ab109028. 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

    ab226049 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    Western blot - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    All lanes : Anti-HP1 alpha antibody [EPR5777] - Heterochromatin marker (ab109028) at 1/1000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate
    Lane 3 : Mouse brain lysate
    Lane 4 : Rat brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 22 kDa



    We are unsure how to define the extra bands.

    Purified ab109028 at 1/30 dilution (2µg) immunoprecipitating HP1 alpha in MCF7 whole cell lysate.

  • Immunoprecipitation - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    Immunoprecipitation - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)

    Purified ab109028 at 1/30 dilution (2µg) immunoprecipitating HP1 alpha in MCF7 whole cell lysate.
    Lane 1 (input): MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab109028 + MCF7 whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109028 in MCF7 whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 22 kDa
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109028).

  • Immunocytochemistry - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    Immunocytochemistry - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    This data was developed using ab109028, the same antibody clone in a different buffer formulation.
    Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HP1 alpha with Purified ab109028 at 1/250 dilution (1.22 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Flow Cytometry - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    Flow Cytometry - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    This data was developed using ab109028, the same antibody clone in a different buffer formulation.
    Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling HP1 alpha with Purified ab109028 at 1/30 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    This data was developed using ab109028, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labeling HP1 alpha with Purified ab109028 at 1/1000 dilution (0.31 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunocytochemistry - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    Immunocytochemistry - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)

    ab109028 staining HP1 alpha in wild-type HAP1 cells (top panel) and HP1 alpha 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 ab109028 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.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109028).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    This data was developed using ab109028, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling HP1 alpha with Purified ab109028 at 1/1000 dilution (0.31 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    This data was developed using ab109028, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat pancreas tissue sections labeling HP1 alpha with Purified ab109028 at 1/1000 dilution (0.31 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)
    Anti-HP1 alpha antibody [EPR5777] - BSA and Azide free (ab226049)

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