Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6322] to CBFb - BSA and Azide free
  • Suitable for: ICC/IF, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-CBFb antibody [EPR6322] - BSA and Azide free
    See all CBFb primary antibodies

  • Description

    Rabbit monoclonal [EPR6322] to CBFb - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, WBmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: A431, Raji, HeLa, K562, Jurkat, and Raw264.7 cell lysates. Mouse and rat spleen and thymus tissue lysates; ICC/IF: Jurkat cells; Flow Cyt: K562 cells.

  • General notes

    Ab248572 is the carrier-free version of ab133600. 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

    ab248572 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.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Dissociation constant (KD)

    KD = 7.20 x 10 -11 M
    Learn more about KD

  • Storage buffer

    pH: 7.2
    Constituent: PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR6322

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)
    Western blot - Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)
    All lanes : Anti-CBFb antibody [EPR6322] (ab133600) at 1/1000 dilution

    Lane 1 : Wild-type A431 cell lysate
    Lane 2 : CBFB knockout A431 cell lysate
    Lane 3 : K562 cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 22 kDa
    Observed band size: 24 kDa
    why is the actual band size different from the predicted?


    Exposure time: 150 seconds


    ab133600 was shown to react with CBFb in wild-type A431 cells in western blot. Loss of signal was observed when CBFB knockout sample was used. Membranes were blocked in 2 % BSA in TBS-T (0.1 % Tween®) before incubation with ab133600 overnight at 4°C at a 1 in 1000 dilution and ab184095 (Mouse Anti-GAPDH antibody [mAbcam 9484] - Alexa Fluor® 680) at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1/5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)
    Immunocytochemistry/ Immunofluorescence - Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)

    Immunocytochemistry/Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CBFb with purified ab133600 at 1/100 dilution (10 µg/mL). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133600).

  • Flow Cytometry - Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)
    Flow Cytometry - Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)

    Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells, labeling CBFb with purified ab133600 at 1/100 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluo® 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133600).

  • Western blot - Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)
    Western blot - Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)
    All lanes : Anti-CBFb antibody [EPR6322] (ab133600) at 1/1000 dilution

    Lane 1 : Wild-type A431 cell lysate
    Lane 2 : CBFB knockout A431 cell lysate
    Lane 3 : K562 cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 22 kDa
    Observed band size: 22 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab133600).

    Lanes 1 - 4: Merged signal (red and green). Green - ab133600 observed at 22 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab133600 was shown to react with CBFb in wild-type A431 cells in western blot with loss of signal observed in CBFB knockout sample. Wild-type and CBFB knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 2% BSA in TBS-T (0.1% Tween®) before incubation with ab133600 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution 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.

  • Western blot - Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)
    Western blot - Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)
    All lanes : Anti-CBFb antibody [EPR6322] (ab133600) at 1/1000 dilution (Purified)

    Lane 1 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
    Lane 4 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
    Lane 5 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 6 : Mouse spleen lysate
    Lane 7 : Mouse thymus lysate
    Lane 8 : Rat spleen lysate
    Lane 9 : Rat thymus 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
    Observed band size: 22 kDa



    Blocking/Diluting buffer: 5% NFDM/TBST

    Loading Control: Rabbit monoclonal [EPR16891] to GAPDH (ab181602)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133600).

  • Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)
    Anti-CBFb antibody [EPR6322] - BSA and Azide free (ab248572)

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