Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-GBA antibody [EPR5143(3)] (ab128879)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5143(3)] to GBA
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Rat, Human

Overview

  • Product name

    Anti-GBA antibody [EPR5143(3)]
    See all GBA primary antibodies

  • Description

    Rabbit monoclonal [EPR5143(3)] to GBA

  • Host species

    Rabbit

  • Specificity

    The lab re-tested the antibody in mouse samples without obtaining satisfactory results (tissue specific positive and negative results), therefore we are not able to guarantee the antibody in this species. Please contact our Scientific Support if you have any feedback in mouse.

  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    WB
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human GBA aa 500-600 (internal sequence). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa, Hap1, 293T, Saos-2, C6 and U87-MG cell lysates. IHC-P: Human kidney tissue.

  • General notes

    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

Images

  • Western blot - Anti-GBA antibody [EPR5143(3)] (ab128879)
    Western blot - Anti-GBA antibody [EPR5143(3)] (ab128879)
    All lanes : Anti-GBA antibody [EPR5143(3)] (ab128879) at 1/1000 dilution

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

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 60 kDa
    Observed band size: 60 kDa



    Lanes 1- 2: Merged signal (red and green). Green - ab128879 observed at 60 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab128879 was shown to react with GBA in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265038 (knockout cell lysate ab256929) was used. Wild-type HeLa and GBA 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. ab128879 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-GBA antibody [EPR5143(3)] (ab128879)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBA antibody [EPR5143(3)] (ab128879)

    ab128879 staining GBA in Human kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/200). An undiluted HRP-conjugated anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin.

  • Western blot - Anti-GBA antibody [EPR5143(3)] (ab128879)
    Western blot - Anti-GBA antibody [EPR5143(3)] (ab128879)
    All lanes : Anti-GBA antibody [EPR5143(3)] (ab128879) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : GBA knockout HAP1 whole cell lysate
    Lane 3 : MCF7 whole cell lysate
    Lane 4 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 60 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab128879 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab128879 was shown to specifically react with GBA in wild-type HAP1 cells as well as additional cross reactive bands. No bands were observed when GBA knockout samples were used. Wild-type and GBA knockout samples were subjected to SDS-PAGE. Ab128879 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with 800CW Goat anti-Rabbit and 680CW Goat anti-Mouse secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-GBA antibody [EPR5143(3)] (ab128879)
    Western blot - Anti-GBA antibody [EPR5143(3)] (ab128879)
    Lane 1 : Anti-GBA antibody [EPR5143(3)] (ab128879) at 1/10000 dilution
    Lanes 2-4 : Anti-GBA antibody [EPR5143(3)] (ab128879) at 1/50000 dilution

    All lanes : C6 (Rat glial tumor glial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/100000 dilution

    Predicted band size: 60 kDa
    Observed band size: 60 kDa


    Exposure time: 180 seconds


    Blocking and diluting buffers:

    Lane 1 and 2: 5% NFDM/TBST
    Lane 3 and 4: 2% BSA/TBST

    We suggest using 2% BSA/TBST as blocking buffer and antibody diluting buffer if you cannot obtain strong band in some samples.

  • Western blot - Anti-GBA antibody [EPR5143(3)] (ab128879)
    Western blot - Anti-GBA antibody [EPR5143(3)] (ab128879)
    All lanes : Anti-GBA antibody [EPR5143(3)] (ab128879) at 1/4000 dilution

    Lane 1 : Saos-2 Cell Lysate
    Lane 2 : U87-MG Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), HRP- conjugated at 1/1000 dilution

    Predicted band size: 60 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBA antibody [EPR5143(3)] (ab128879)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBA antibody [EPR5143(3)] (ab128879)

    ab128879, unpurified, at a 1/100 dilution, staining GBA in paraffin embedded Human kidney tissue by Immunohistochemistry.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • OI-RD Scanning - Anti-GBA antibody [EPR5143(3)] (ab128879)
    OI-RD Scanning - Anti-GBA antibody [EPR5143(3)] (ab128879)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Western blot - Anti-GBA antibody [EPR5143(3)] (ab128879)
    Western blot - Anti-GBA antibody [EPR5143(3)] (ab128879)
    All lanes : Anti-GBA antibody [EPR5143(3)] (ab128879) at 1/1000 dilution (unpurified)

    Lane 1 : 293T cell lysate
    Lane 2 : Saos-2 cell lysate
    Lane 3 : U87-MG cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 60 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBA antibody [EPR5143(3)] (ab128879)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBA antibody [EPR5143(3)] (ab128879)

    ab128879, unpurified, at a 1/100 dilution, staining GBA in paraffin embedded Human thyroid carcinoma tissue by Immunohistochemistry.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Anti-GBA antibody [EPR5143(3)] (ab128879)
    Anti-GBA antibody [EPR5143(3)] (ab128879)

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