Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-GBA antibody [EPR5142] - BSA and Azide free (ab215261)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5142] to GBA - BSA and Azide free
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-GBA antibody [EPR5142] - BSA and Azide free
    See all GBA primary antibodies

  • Description

    Rabbit monoclonal [EPR5142] to GBA - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WBmore details
    Unsuitable for: Flow Cyt,ICC or IP

  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat

  • Immunogen

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

  • Positive control

    • IHC-P: Human lung cancer and colon tissue; WB: HeLa, HAP1, MCF7 and HepG2 cell lysates.

  • General notes

    Ab215261 is the carrier-free version of ab125065. 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

    ab215261 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Images

  • Western blot - Anti-GBA antibody [EPR5142] - BSA and Azide free (ab215261)
    Western blot - Anti-GBA antibody [EPR5142] - BSA and Azide free (ab215261)
    All lanes : Anti-GBA antibody [EPR5142] (ab125065) 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: 70 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 (ab125065).

      Lanes 1- 2: Merged signal (red and green). Green - ab125065 observed at 70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab125065 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. ab125065 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 [EPR5142] - BSA and Azide free (ab215261)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBA antibody [EPR5142] - BSA and Azide free (ab215261)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling GBA with Purified ab125065 at 1:50 dilution (2.1 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125065). 

     

  • Western blot - Anti-GBA antibody [EPR5142] - BSA and Azide free (ab215261)
    Western blot - Anti-GBA antibody [EPR5142] - BSA and Azide free (ab215261)
    All lanes : Anti-GBA antibody [EPR5142] (ab125065) 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 40 µg per lane.

    Predicted band size: 60 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab125065 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab125065 was shown to specifically recognize 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. Ab125065 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing tissue culture supernatant, tris glycine, BSA, glycerol and sodium azide (ab125065).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBA antibody [EPR5142] - BSA and Azide free (ab215261)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GBA antibody [EPR5142] - BSA and Azide free (ab215261)

    ab125065, at a 1/50 dilution, staining GBA in paraffin-embedded Human colon tissue by immunohistochemistry.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab125065). 

    This image was generated using the unpurified version of the product. 

    Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-GBA antibody [EPR5142] - BSA and Azide free (ab215261)
    Anti-GBA antibody [EPR5142] - BSA and Azide free (ab215261)

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