Anti-MARCO antibody [EPR22944-64]

7 product images

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  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 (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 (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.

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  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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab239369 in mouse spleen lysate.

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

  Exposure time: 8 seconds.

  All lanes: Immunoprecipitation - Anti-MARCO antibody [EPR22944-64] (ab239369)

  Predicted band size: 53 kDa

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  Western blot - Anti-MARCO antibody [EPR22944-64] (ab239369)

  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).

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

  Lane 1: Untreated J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), whole cell lysate at 20 µg

  Lane 2: J774A.1 treated with 1ug/ml Lipopolysaccharides (LPS) for 24 hours, whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 53 kDa

  Observed band size: 55-75 kDa

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  Western blot - Anti-MARCO antibody [EPR22944-64] (ab239369)

  Blocking and Dilution Buffer and concentration: 5% NFDM/TBSTThe expression of MARCO/Macrophage receptor with collagenous structure is induced by Lipopolysaccharides (LPS) treatment (PMID: 9916718) 3 seconds

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

  Lane 1: Mouse spleen lysate at 20 µg

  Lane 2: Mouse spleen lysate treated with Peptide:N-glycosidase F (PNGase F) at 20 µg

  Secondary

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

  Predicted band size: 53 kDa

  Observed band size: 53 kDa, 75 kDa

  Exposure time: 15s

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  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 (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 (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.

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  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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).

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  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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.