Recombinant
RabMAb

Recombinant Anti-Granulin antibody [EPR18539-59] (ab187070)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18539-59] to Granulin
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt
  • Reacts with: Mouse

Overview

  • Product name

    Anti-Granulin antibody [EPR18539-59]
    See all Granulin primary antibodies

  • Description

    Rabbit monoclonal [EPR18539-59] to Granulin

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

    Recombinant fragment corresponding to Mouse Granulin aa 350 to the C-terminus.
    Database link: P28798

  • Positive control

    • WB: Mouse spleen tissue lysate; J774A.1 whole cell lysate. IHC-P: Mouse lung tissue. ICC/IF: J774A.1 cells. Flow Cytometry: J774A.1 cells. IP: J774A.1 whole cell lysate.

  • General notes

    ab187070 is also recommended for human and rat species for IHC.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab187070 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 1/1000. Predicted molecular weight: 63 kDa.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/30.
Flow Cyt 1/1000.

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

    • Western blot - Anti-Granulin antibody [EPR18539-59] (ab187070)
      Western blot - Anti-Granulin antibody [EPR18539-59] (ab187070)
      Lane 1 : Anti-Granulin antibody [EPR18539-59] (ab187070) at 1/1000 dilution
      Lane 2 : Anti-Granulin antibody [EPR18539-59] (ab187070) at 1/5000 dilution

      Lane 1 : J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage cell line) whole cell lysate
      Lane 2 : Mouse spleen tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Developed using the ECL technique.

      Predicted band size: 63 kDa
      Observed band size: 88 kDa
      why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times.

      Lane 1: 3 seconds.

      Lane 2: 10 seconds.

      The molecular mass observed is consistent with what has been described in the literature (PMID: 23972823). Granulin (Progranulin, PGRN) is a 593-amino acid, 68.5-kDa cysteine-rich protein that is typically secreted in a highly glycosylated 88-kDa form.

    • Immunocytochemistry/ Immunofluorescence - Anti-Granulin antibody [EPR18539-59] (ab187070)
      Immunocytochemistry/ Immunofluorescence - Anti-Granulin antibody [EPR18539-59] (ab187070)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte macrophage cell line) cells labeling Granulin with ab187070 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on J774A.1 cell line.

      The nuclear counter statin is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

      Negative control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Flow Cytometry - Anti-Granulin antibody [EPR18539-59] (ab187070)
      Flow Cytometry - Anti-Granulin antibody [EPR18539-59] (ab187070)

      Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) cell line labeling Granulin with ab187070 at 1/1000 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue).

      Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

    • Immunoprecipitation - Anti-Granulin antibody [EPR18539-59] (ab187070)
      Immunoprecipitation - Anti-Granulin antibody [EPR18539-59] (ab187070)

      Granulin was immunoprecipitated from 0.35 mg of J774A.1 (mouse reticulum cell sarcoma monocyte macrophage cell line) whole cell lysate with ab187070 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab187070 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

      Lane 1: J774A.1 lysate 10 μg (Input).

      Lane 2: ab187070 IP in J774A.1 lysate (+).

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab187070 in J744A.1 whole cell lysate (-).

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 3 minutes.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granulin antibody [EPR18539-59] (ab187070)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granulin antibody [EPR18539-59] (ab187070)

      Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling Granulin with ab187070 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Sporadic cytoplasmic staining in mouse lung (PMID: 23959579) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References

    Publishing research using ab187070? Please let us know so that we can cite the reference in this datasheet.

    ab187070 has not yet been referenced specifically in any publications.

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