重组Anti-MARCO抗体[EPR22944-64]

• IHC-P

Unknown

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.

• ICC/IF

Unknown

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.

• IHC-P

Unknown

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.

• IHC-Fr

Unknown

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).

• IP

Unknown

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.

All lanes:

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

Predicted band size: 53 kDa

false

• WB

Supplier Data

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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 53 kDa

Observed band size: 55-75 kDa

false

• WB

Lab

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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa,75 kDa

false

Exposure time: 15s