Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15864] to Granulin - Low endotoxin, Azide free
  • Suitable for: WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free
    See all Granulin primary antibodies

  • Description

    Rabbit monoclonal [EPR15864] to Granulin - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: HepG2, HAP1, A431 and HEK293T whole cell lysate; Human fetal kidney lysate; Human GRN full-length recombinant protein with GST-tag at N-terminal. IHC-P: Human liver, placenta, breast cancer and gastric cancer tissues. IP: HeLa whole cell lysate.

  • General notes

    ab225532 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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.

Properties

Images

  • Western blot - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)
    Western blot - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)
    All lanes : Anti-Granulin antibody [EPR15864] (ab208777) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : GRN knockout HEK293T cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 64 kDa
    Observed band size: 74 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 (ab208777).

    Lanes 1-4: Merged signal (red and green). Green - ab208777 observed at 74 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab208777 Anti-Granulin antibody [EPR15864] was shown to specifically react with Granulin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266738 (knockout cell lysate ab257235) was used. Wild-type and Granulin knockout samples were subjected to SDS-PAGE. ab208777 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 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-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)

    Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Granulin with ab208777 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Kupffer cells of Human liver is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208777).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)

    Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling Granulin with ab208777 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on trophoblastic cells of Human placenta is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208777).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)

    Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Granulin with ab208777 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human breast cancer is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208777).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)

    This IHC data was generated using the same anti-Granulin antibody clone [EPR15864] in a different buffer formulation (cat# ab208777).

    Immunohistochemical analysis of paraffin-embedded Human gastric cancer tissue labeling Granulin with ab208777 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on Human gastric cancer is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)
    Western blot - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)
    All lanes : Anti-Granulin antibody [EPR15864] (ab208777) at 1/2000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : GRN (Granulin) knockout HAP1 whole cell lysate
    Lane 3 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 64 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab208777 observed at 64 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab208777 was shown to specifically react with Granulin in wild-type HAP1 cells as signal was lost in GRN (Granulin) knockout cells. Wild-type and GRN (Granulin) knockout samples were subjected to SDS-PAGE. Ab208777 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208777).

  • Immunoprecipitation - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)
    Immunoprecipitation - Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)

    Granulin was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab208777 at 1/70 dilution.

    Western blot was performed from the immunoprecipitate using ab208777 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate, 10µg (Input).

    Lane 2: ab208777 IP in HeLa whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal -Isotype Control (ab172730) instead of ab208777 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208777).

  • Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)
    Anti-Granulin antibody [EPR15864] - Low endotoxin, Azide free (ab225532)

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