RecombinantRabMAb

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

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

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1179Y] to M-CSF
  • Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt
  • Reacts with: Human

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

     

     

    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.

    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.

    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

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.

    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 (16)

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

    ab52864 has been referenced in 16 publications.

    • 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). 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). PubMed: 30580386
    • Morizawa Y  et al. Colony-stimulating factors detected in tumor cells and voided urine are potential prognostic markers for patients with muscle-invasive bladder cancer undergoing radical cystectomy. Res Rep Urol 10:103-111 (2018). PubMed: 30288389
    • Wang H  et al. Interactions between colon cancer cells and tumor-infiltrated macrophages depending on cancer cell-derived colony stimulating factor 1. Oncoimmunology 5:e1122157 (2016). PubMed: 27141406
    • Yang L  et al. Increased expression of colony stimulating factor-1 is a predictor of poor prognosis in patients with clear-cell renal cell carcinoma. BMC Cancer 15:67 (2015). IHC-P ; Human . PubMed: 25886010
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    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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