Anti-Macrophage antibody [RM0029-11H3] (ab56297)

Overview

  • Product nameAnti-Macrophage antibody [RM0029-11H3]
    See all Macrophage primary antibodies
  • Description
    Rat monoclonal [RM0029-11H3] to Macrophage
  • Tested applicationsSuitable for: ICC/IF, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Isolated mouse peritoneal macrophages

  • Positive control
    • Mouse kidney tissue. IF/ICC: RAW246.7

Properties

Applications

Our Abpromise guarantee covers the use of ab56297 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
ICC/IF Use a concentration of 10 µg/ml.
IP 1/100.
Flow Cyt Use 2µg for 106 cells. ab18450-Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
IHC-P Use at an assay dependent concentration. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.

Target

  • RelevanceMacrophages comprise of many forms of mononuclear phagocytes found in tissues. Mononuclear phagocytes arise from hematopoietic stem cells in the bone marrow. After passing through the monoblast and promonocyte states of the monocyte stage, they enter the blood, where they circulate for about 40 hours. They then enter tissues and increase in size, phagocytic activity, and lysosomal enzyme content becomming macrophages. Among the functions of macrophages are nonspecific phagocytosis and pinocytosis, specific phagocytosis of opsonized microorganisms mediated by Fc receptors and complement receptors, killing of ingested microorganisms, digestion and presentation of antigens to T and B lymphocytes, and secretion of a large number of diverse products, including many enzymes including lysozyme and collagenases, several complement components and coagulation factors, some prostaglandins and leukotrienes, and many regulatory molecules (Interferon, Interleukin 1). Among cells that are now recognised as macrophages are histiocytes, Kupffer cells, osteoclasts, microglial cells, synovial type A cells, interdigitating cells, and Langerhans cells (in normal tissues) and epithelioid cells and Langerhans-type and foreign-body-type multinucleated giant cells (in inflamed tissues).
  • Alternative names
    • macrophages antibody

Anti-Macrophage antibody [RM0029-11H3] images

  • ICC/IF image of ab56297 stained RAW246.7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab56297, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96887, DyLight® 488 goat anti-rat IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM

  • Bouin’s solution fixed and paraffin embedded mouse kidney section from anti-GBM model was subjected to immunohistochemistry staining (ABC) of Macrophage using ab56297.Bouin’s solution fixed and paraffin embedded mouse kidney section from anti-GBM model was subjected to immunohistochemistry staining (ABC) of Macrophage using ab56297.
  • Overlay histogram showing RAW 264.7 cells stained with ab56297 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab56297, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rat IgG2a, kappa monoclonal [aRTK2758] (ab18450, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in RAW 264.7 cells fixed with 80% methanol/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Immunohistochemical analysis of murine tumour tissue, staining Macrophage with ab56297. Antigen retrieval was performed under high pressure in 10 mM EDTA buffer (pH 8.0) before incubation with primary antibody.

References for Anti-Macrophage antibody [RM0029-11H3] (ab56297)

This product has been referenced in:
  • Mohamed R  et al. Urinary semaphorin 3A correlates with diabetic proteinuria and mediates diabetic nephropathy and associated inflammation in mice. J Mol Med (Berl) N/A:N/A (2014). Mouse . Read more (PubMed: 25249008) »
  • Lucas AR  et al. Myxomavirus anti-inflammatory chemokine binding protein reduces the increased plaque growth induced by chronic Porphyromonas gingivalis oral infection after balloon angioplasty aortic injury in mice. PLoS One 9:e111353 (2014). IHC ; Mouse . Read more (PubMed: 25354050) »

See all 9 Publications for this product

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The antibody is covered under our Abpromise for six months and is guaranteed to work in IHC-P on mouse samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either s...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (brain tumor)
Specification brain tumor
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Blocking step (agent) for 30 minute(s) · Concentration: 0.5% · Temperature: 25°C
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Submitted Apr 22 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"