Recombinant
RabMAb

Recombinant Anti-M-CSF antibody [EP1179Y] (ab52864)

Submit a review Q&A (1)References (16)

Overview

  • Product name

    Anti-M-CSF antibody [EP1179Y]
    See all M-CSF primary antibodies

  • Description

    Rabbit monoclonal [EP1179Y] to M-CSF

  • Host species

    Rabbit

  • Specificity

    This antibody fails to detect endogenous natural samples in WB.

  • Tested applications

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

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Sheep, Cow, Chimpanzee

  • Immunogen

    Synthetic peptide within Human M-CSF aa 50-150. The exact sequence is proprietary.
    Database link: P09603

  • Positive control

    • WB: Recombinant MCSF IHC-P: Human tonsil tissue.

  • General notes

     

     

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52864 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 1/2000. Predicted molecular weight: 60 kDa.

This antibody fails to detect endogenous natural samples in WB.
IP 1/40.

For unpurified use at 1/50 dilution. 
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/50 - 1/100 dilution.
ICC/IF 1/100.

For unpurified use at 1/250 - 1/500 dilution.
Flow Cyt 1/80.

Target

  • Function

    Granulocyte/macrophage colony-stimulating factors are cytokines that act in hematopoiesis by controlling the production, differentiation, and function of 2 related white cell populations of the blood, the granulocytes and the monocytes-macrophages. CSF-1 induces cells of the monocyte/macrophage lineage. It plays a role in immunological defenses, bone metabolism, lipoproteins clearance, fertility and pregnancy.

  • Post-translational
    modifications

    Glycosylation and proteolytic cleavage yield different soluble forms. A high molecular weight soluble form is a proteoglycan containing chondroitin sulfate.
    Isoform 1 is N- and O-glycosylated. Isoform 3 is N-glycosylated.

  • Cellular localization

    Cell membrane and Secreted > extracellular space.
  • Information by UniProt

  • Database links

    • Alternative names

      • Colony stimulating factor 1 (macrophage) antibody
      • Colony stimulating factor 1 antibody
      • Colony stimulating factor macrophage specific antibody
      • CSF 1 antibody
      • CSF-1 antibody
      • CSF1 antibody
      • CSF1_HUMAN antibody
      • Csfm antibody
      • Lanimostim antibody
      • M CSF antibody
      • M-CSF antibody
      • Macrophage Colony Stimulating Factor 1 antibody
      • Macrophage colony stimulating factor antibody
      • MCSF antibody
      • MGC31930 antibody
      • Processed macrophage colony-stimulating factor 1 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M-CSF antibody [EP1179Y] (ab52864)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M-CSF antibody [EP1179Y] (ab52864)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling M-CSF with Purified ab52864 at 1:500 dilution (1.52 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0)

    • Immunocytochemistry/ Immunofluorescence - Anti-M-CSF antibody [EP1179Y] (ab52864)
      Immunocytochemistry/ Immunofluorescence - Anti-M-CSF antibody [EP1179Y] (ab52864)

      Immunocytochemistry/ Immunofluorescence analysis of THP-1 (Human monocytic leukemia monocyte) cells labeling M-CSF with Purified ab52864 at 1:100 (7.6 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Western blot - Anti-M-CSF antibody [EP1179Y] (ab52864)
      Western blot - Anti-M-CSF antibody [EP1179Y] (ab52864)
      Anti-M-CSF antibody [EP1179Y] (ab52864) at 1/2000 dilution (Purified) + Human M-CSF (aa 33-190) recombinant protein at 0.015 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 60 kDa
      Observed band size: 19 kDa
      why is the actual band size different from the predicted?



      Blocking buffer: 5% NFDM/TBST.

    • Flow Cytometry - Anti-M-CSF antibody [EP1179Y] (ab52864)
      Flow Cytometry - Anti-M-CSF antibody [EP1179Y] (ab52864)

      Flow Cytometry analysis of THP-1 (Human monocytic leukemia monocyte) cells labeling M-CSF with purified ab52864 at 1:80 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M-CSF antibody [EP1179Y] (ab52864)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-M-CSF antibody [EP1179Y] (ab52864)

      Formalin-fixed, paraffin-embedded human tonsil tissue stained for M-CSF with unpurified ab52864  (1/50 dilution) in immunohistochemical analysis.

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

    • Western blot - Anti-M-CSF antibody [EP1179Y] (ab52864)
      Western blot - Anti-M-CSF antibody [EP1179Y] (ab52864)
      Anti-M-CSF antibody [EP1179Y] (ab52864) at 1/50000 dilution (unpurified) + recombinant MCSF at 0.005 µg

      Secondary
      Goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 60 kDa
      Observed band size: 20 kDa why is the actual band size different from the predicted?

    • Immunoprecipitation - Anti-M-CSF antibody [EP1179Y] (ab52864)
      Immunoprecipitation - Anti-M-CSF antibody [EP1179Y] (ab52864)

      ab52864 (purified) at 1:40 dilution (2µg) immunoprecipitating M-CSF in THP-1 whole cell lysate.
      Lane 1 (input): THP-1 (Human monocytic leukemia monocyte) whole cell lysate 10µg
      Lane 2 (+): ab52864 & THP-1 whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52864 in THP-1 whole cell lysate
      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    References

    This product has been referenced in:

    • Baghdadi M  et al. High co-expression of IL-34 and M-CSF correlates with tumor progression and poor survival in lung cancers. Sci Rep 8:418 (2018). Read more (PubMed: 29323162) »
    • de Vries WM  et al. M-CSF and IL-34 expression as indicators for growth in sporadic vestibular schwannoma. Virchows Arch N/A:N/A (2018). Read more (PubMed: 30580386) »
    See all 16 Publications for this product

    Publishing research using ab52864? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    Submit a review Submit a question

    Question

    I've been using your antibody against M-CSF (ab52864). It says it is a 60kDa protein but is observed at 20kDa. When tested in my samples, this protein is coming up at around 45kDa. Have you seen this before? It is a very clean blot with only one specific band present so I do believe the antibody is very specific but just wondering if you have any advice in this case?


     



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    Answer

    I can confirm to you that it is reported in literature that M-CSF can also be detected in a 45 kDa form (see links below)

    https://www.ncbi.nlm.nih.gov/pubmed/2179407

    https://www.researchgate.net/publication/14333823_Macrophage_colony_stimulating_factor_involvement_in_uremic_patients  (in the introduction section).

    The size of the band is dependent on whether you are detecting the circulating or membrane bound form.

    I hope you found this information helpful and I am happy to answer any further questions or concerns you have.  The antibody is covered by the Abpromise guarantee.

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