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RecombinantRabMAb

Recombinant Anti-MARCO antibody [EPR22944-64] (ab239369)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MARCO antibody [EPR22944-64] (ab239369)
  • Western blot - Anti-MARCO antibody [EPR22944-64] (ab239369)
  • Western blot - Anti-MARCO antibody [EPR22944-64] (ab239369)
  • Immunoprecipitation - Anti-MARCO antibody [EPR22944-64] (ab239369)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MARCO antibody [EPR22944-64] (ab239369)
  • Immunohistochemistry (Frozen sections) - Anti-MARCO antibody [EPR22944-64] (ab239369)
  • Immunocytochemistry/ Immunofluorescence - Anti-MARCO antibody [EPR22944-64] (ab239369)
  • Anti-MARCO antibody [EPR22944-64] (ab239369)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22944-64] to MARCO
  • Suitable for: IP, IHC-Fr, IHC-P, ICC/IF, WB
  • Reacts with: Mouse

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

Overview

  • Product name

    Anti-MARCO antibody [EPR22944-64]
    See all MARCO primary antibodies

  • Description

    Rabbit monoclonal [EPR22944-64] to MARCO

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, IHC-Fr, IHC-P, ICC/IF, WBmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: J774A.1 treated with Lipopolysaccharides whole cell lysate. Mouse spleen lysate. Mouse spleen lysate treated with Peptide: N-glycosidase F (PNGase F). IHC-P: Mouse spleen and liver tissue. IHC-Fr: Mouse spleen tissue. ICC/IF: J774A.1 cells treated with Lipopolysaccharides. IP: Mouse spleen lysate.

  • 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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab239369 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
IP
1/30.
IHC-Fr
1/500.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100.
WB
1/1000.
Notes
IP
1/30.
IHC-Fr
1/500.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/100.
WB
1/1000.
Application notes
Is unsuitable for Flow Cyt.

Target

  • Function

    Pattern recognition receptor (PRR). Binds Gram-positive and Gram-negative bacteria.

  • Sequence similarities

    Contains 1 collagen-like domain.
    Contains 1 SRCR domain.

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • AI323439 antibody
      • Ly112 antibody
      • Macrophage receptor MARCO antibody
      • Macrophage receptor with collagenous structure antibody
      • Marco antibody
      • MARCO_HUMAN antibody
      • SCARA2 antibody
      • Scavenger receptor class A member 2 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MARCO antibody [EPR22944-64] (ab239369)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MARCO antibody [EPR22944-64] (ab239369)

      Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling MARCO with ab239369 at 1/500 dilution (1.11 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Kupffer cells of mouse liver is observed. The section was incubated with ab239369 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND®RX instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

    • Western blot - Anti-MARCO antibody [EPR22944-64] (ab239369)
      Western blot - Anti-MARCO antibody [EPR22944-64] (ab239369)
      All lanes : Anti-MARCO antibody [EPR22944-64] (ab239369) at 1/1000 dilution

      Lane 1 : Untreated J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), whole cell lysate
      Lane 2 : J774A.1 treated with 1ug/ml Lipopolysaccharides (LPS) for 24 hours, whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Observed band size: 55-75 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST .

      Exposure time: 3 seconds.

      The expression of MARCO/Macrophage receptor with collagenous structure is induced by Lipopolysaccharides (LPS) treatment (PMID: 9916718).

    • Western blot - Anti-MARCO antibody [EPR22944-64] (ab239369)
      Western blot - Anti-MARCO antibody [EPR22944-64] (ab239369)
      All lanes : Anti-MARCO antibody [EPR22944-64] (ab239369) at 1/1000 dilution

      Lane 1 : Mouse spleen lysate
      Lane 2 : Mouse spleen lysate treated with Peptide:N-glycosidase F (PNGase F)

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Observed band size: 53,75 kDa why is the actual band size different from the predicted?


      Exposure time: 15 seconds


      75 kDa (glycosylated from), 53 kDa (deglycosylated form).

      Blocking and Dilution Buffer and concentration: 5% NFDM/TBST.

      N-glycosylation is crucial for MARCO/Macrophage receptor with collagenous structure intercellular trafficking and surface targeting (PMID: 26892079).

    • Immunoprecipitation - Anti-MARCO antibody [EPR22944-64] (ab239369)
      Immunoprecipitation - Anti-MARCO antibody [EPR22944-64] (ab239369)

      MARCO was immunoprecipitated from 0.35 mg mouse spleen lysate 10µg with ab239369 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab239369 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

      Lane 1: Mouse spleen lysate 10µg.

      Lane 2: ab239369 IP in mouse spleen lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239369 in mouse spleen lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 8 seconds.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MARCO antibody [EPR22944-64] (ab239369)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MARCO antibody [EPR22944-64] (ab239369)

      Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling MARCO with ab239369 at 1/500 dilution (1.11 µg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on macrophages of mouse spleen (PMID: 12874264, 15494482) is observed. The section was incubated with ab239369 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® instrument. Counterstained with hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

    • Immunohistochemistry (Frozen sections) - Anti-MARCO antibody [EPR22944-64] (ab239369)
      Immunohistochemistry (Frozen sections) - Anti-MARCO antibody [EPR22944-64] (ab239369)

      4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen section of mouse spleen tissue labeling MARCO using ab239369 at 1/500 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (green). The nuclear counterstain is DAPI (blue). Positive staining on mouse splenic marginal zone (PMID: 24670793) is observed.

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

    • Immunocytochemistry/ Immunofluorescence - Anti-MARCO antibody [EPR22944-64] (ab239369)
      Immunocytochemistry/ Immunofluorescence - Anti-MARCO antibody [EPR22944-64] (ab239369)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized J744A.1 (Mouse reticulum cell sarcoma monocyte macrophage) cells labeling MARCO with ab239369 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in J744A.1 cells treated with LPS (1 μg/ml) for 24h. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

    • Anti-MARCO antibody [EPR22944-64] (ab239369)
      Anti-MARCO antibody [EPR22944-64] (ab239369)

    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 (0)

    Publishing research using ab239369? Please let us know so that we can cite the reference in this datasheet.

    ab239369 has not yet been referenced specifically in any publications.

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