Recombinant
RabMAb

Recombinant Anti-MARCO/Macrophage receptor with collagenous structure antibody [EPR22944-64] (ab239369)

Overview

  • Product name

    Anti-MARCO/Macrophage receptor with collagenous structure antibody [EPR22944-64]
    See all MARCO/Macrophage receptor with collagenous structure primary antibodies
  • Description

    Rabbit monoclonal [EPR22944-64] to MARCO/Macrophage receptor with collagenous structure
  • 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 within Mouse MARCO/Macrophage receptor with collagenous structure aa 1-100. The exact sequence is proprietary.
    Database link: Q60754

  • 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

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.
  • 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

    • All lanes : Anti-MARCO/Macrophage receptor with collagenous structure antibody [EPR22944-64] (ab239369) at 1/1000 dilution

      Lane 1 : Untreated J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), whole cell lysate 20 µg
      Lane 2 : J774A.1 treated with 1µg/ml Lipopolysaccharides (LPS) for 24 hours, whole cell lysate 20 µg

      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?


      Exposure time: 3 seconds


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

      Blocking buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 seconds.

       

    • All lanes : Anti-MARCO/Macrophage receptor with collagenous structure 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).

    • MARCO/Macrophage receptor with collagenous structure 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.

       

    • Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling MARCO/Macrophage receptor with collagenous structure 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.

    • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling MARCO/Macrophage receptor with collagenous structure 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.

    • 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen section of mouse spleen tissue labeling MARCO/Macrophage receptor with collagenous structure 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).

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized J744A.1 (Mouse reticulum cell sarcoma monocyte macrophage) cells labeling MARCO/Macrophage receptor with collagenous structure 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.

    References

    ab239369 has not yet been referenced specifically in any publications.

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