Validated using a knockout cell line
Recombinant
RabMAb

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

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 aa 100-250. The exact sequence is proprietary.
    Database link: P28799

  • Positive control

    • WB: A431 and 293T 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.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab225532 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration. Detects a band of approximately 74 kDa (predicted molecular weight: 64 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP Use at an assay dependent concentration.

Target

  • Function

    Granulins have possible cytokine-like activity. They may play a role in inflammation, wound repair, and tissue remodeling.
    Granulin-4 promotes proliferation of the epithelial cell line A431 in culture while granulin-3 acts as an antagonist to granulin-4, inhibiting the growth.
  • Tissue specificity

    In myelogenous leukemic cell lines of promonocytic, promyelocytic, and proerythroid lineage, in fibroblasts, and very strongly in epithelial cell lines. Present in inflammatory cells and bone marrow. Highest levels in kidney.
  • Involvement in disease

    Defects in GRN are the cause of ubiquitin-positive frontotemporal dementia (UP-FTD) [MIM:607485]; also known as tau-negative frontotemporal dementia linked to chromosome 17. Frontotemporal dementia (FTD) is the second most common cause of dementia in people under the age of 65 years. It is an autosomal dominant neurodegenerative disease.
  • Sequence similarities

    Belongs to the granulin family.
  • Post-translational
    modifications

    Granulins are disulfide bridged.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Database links

  • Alternative names

    • Acrogranin antibody
    • CLN11 antibody
    • GEP antibody
    • GP88 antibody
    • Granulin A antibody
    • Granulin B antibody
    • Granulin C antibody
    • Granulin D antibody
    • Granulin E antibody
    • Granulin epithelin antibody
    • Granulin F antibody
    • Granulin G antibody
    • Granulin-7 antibody
    • Granulins antibody
    • GRN antibody
    • GRN_HUMAN antibody
    • PC cell derived growth factor antibody
    • PCDGF antibody
    • PEPI antibody
    • PGRN antibody
    • Proepithelin antibody
    • Progranulin antibody
    see all

Images

  • 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).

  • 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.

  • 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.

  • 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.

  • 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).

  • 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.

References

ab225532 has not yet been referenced specifically in any publications.

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