All lanes : Anti-MARCO antibody [EPR22944-66] (ab259264) 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: 3 minutes

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

This blot was developed using a higher sensitivity ECL substrate.  N-glycosylation is crucial for MARCO/Macrophage receptor with collagenous structure intercellular trafficking and surface targeting (PMID: 26892079).

All lanes : Anti-MARCO antibody [EPR22944-66] (ab259264) 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?


Exposure time: 6 seconds

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).

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling MARCO with ab259264 at 1/1000 dilution (0.501 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. The section was incubated with ab259264 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 with ab259264 at 1/1000 dilution (0.501 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). The section was incubated with ab259264 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.

4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen section of mouse spleen tissue labeling MARCO using ab259264 at 1/500 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (green). The nuclear counter stain is DAPI (blue). Positive staining on mouse splenic marginal zone (PMID: 24670793).

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% TritonX-100 permeabilized J744A.1 (Mouse reticulum cell sarcoma monocyte macrophage) cells labelling MARCO with ab259264 at 1/50 (10 ug/ml) dilution, followed by Ab259264 anti-MARCO ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 (2 ug/ml) dilution (Green). UNABLE TO ENCODE THE VARIABLE PLEASE DO MANUALLY is observed. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor&~153;,594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is Ab259264 anti-MARCO ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 ug/ml) dilution.