Anti-DAP12 antibody [EPR24244-76]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-DAP12 antibody [EPR24244-76] (AB283679)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse blood cells cells labelling DAP12 with ab283679 at 1/50 dilution (1ug) (Right) compared with a Rabbit monoclonal IgG (ab172730) (Left) isotype control. A Goat F(ab')2 Anti-Rabbit IgG(DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DAP12 antibody [EPR24244-76] (AB283679)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labelling DAP12 with ab283679 at 1/2000 (0.232 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on kupffer cells of rat liver. The section was incubated with ab283679 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DAP12 antibody [EPR24244-76] (AB283679)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labelling DAP12 with ab283679 at 1/2000 (0.232 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat lung. The section was incubated with ab283679 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DAP12 antibody [EPR24244-76] (AB283679)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling DAP12 with ab283679 at 1/2000 (0.232 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse spleen. The section was incubated with ab283679 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DAP12 antibody [EPR24244-76] (AB283679)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling DAP12 with ab283679 at 1/2000 (0.232 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on kupffer cells of mouse liver. The section was incubated with ab283679 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DAP12 antibody [EPR24244-76] (AB283679)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized RAW264.7 cells labelling DAP12 with ab283679 at 1/50 (9.28 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic and membranous staining in RAW 264.7 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Negative control : M1 (PMID : 11034384).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-DAP12 antibody [EPR24244-76] (AB283679)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized M1 (mouse myeloid leukemia myeloblast) (Left), RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (Right) cells labelling DAP12 with ab283679 at 1/500 dilution (0.1ug)(Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab')2 Anti-Rabbit IgG(DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

Negative control : M1 (PMID : 11034384).

• IP

Supplier Data

Immunoprecipitation - Anti-DAP12 antibody [EPR24244-76] (AB283679)

DAP12 was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 ug with ab283679 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283679 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW264.7 whole cell lysate 10 ug

Lane 2 : ab283679 IP in RAW264.7 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283679 in RAW264.7 whole cell lysate

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

Exposure time : 5.5 seconds

All lanes:

Immunoprecipitation - Anti-DAP12 antibody [EPR24244-76] (ab283679)

Predicted band size: 12 kDa

false

• WB

Lab

Western blot - Anti-DAP12 antibody [EPR24244-76] (AB283679)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : M1 (PMID : 11034384).

Exposure time : 114 seconds

All lanes:

Western blot - Anti-DAP12 antibody [EPR24244-76] (ab283679) at 1/1000 dilution

Lane 1:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 2:

N9 (mouse microglia), whole cell lysate at 20 µg

Lane 3:

M1 (mouse myeloid leukemia myeloblast), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 12 kDa

false

• WB

Lab

Western blot - Anti-DAP12 antibody [EPR24244-76] (AB283679)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 3 minutes

All lanes:

Western blot - Anti-DAP12 antibody [EPR24244-76] (ab283679) at 1/1000 dilution

All lanes:

Rat spleen tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 12 kDa

Observed band size: 12 kDa

false