Anti-DAP12 antibody [EPR29151-41] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DAP12 antibody [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling DAP12 with ab323318 at 1/2000 (0.269 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of human colon carcinoma.
The section was incubated with ab323318 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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 [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling DAP12 with ab323318 at 1/2000 (0.269 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen (PMID : 9490415).
The section was incubated with ab323318 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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 [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling DAP12 with ab323318 at 1/2000 (0.269 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on microglia of human cerebrum (PMID : 30124174).
The section was incubated with ab323318 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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 [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling DAP12 with ab323318 at 1/2000 (0.269 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on macrophages of human lung.
The section was incubated with ab323318 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DAP12 antibody [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labelling DAP12 with ab323318 at 1/1000 (0.538 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing increased cytoplasmic staining in THP-1 cell line treated with PMA (100 ng/ml) for 24 hours (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ 200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DAP12 antibody [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling DAP12 with ab323318 at 1/2000 (0.269 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : weak staining on human skeletal muscle.
The section was incubated with ab323318 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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 [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human non-Hodgkin's lymphoma tissue labeling DAP12 with ab323318 at 1/2000 (0.269 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human non-Hodgkin's lymphoma.
The section was incubated with ab323318 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Slides were incubated with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-DAP12 antibody [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

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

Negative control : Jurkat (PMID : 9490415).

The identity of the bands higher 75 kDa are unknown.

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-DAP12 antibody [EPR29151-41] (<a href='/en-us/products/primary-antibodies/dap12-antibody-epr29151-41-ab323318'>ab323318</a>) at 1/1000 dilution

Lane 1:

No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 11 kDa,12 kDa,36 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-DAP12 antibody [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

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

Low expression : skeletal muscle colon testis (PMID : 9490415).

The identity of the bands higher 25 kDa are unknown.

Samples are non-boiled as boiling may cause protein aggregation.

The lanes 2-6 were developed using a high-sensitivity ECL substrate allowing for the detection of proteins in the mid-femtogram range.

In Western blot Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-DAP12 antibody [EPR29151-41] (<a href='/en-us/products/primary-antibodies/dap12-antibody-epr29151-41-ab323318'>ab323318</a>) at 1/1000 dilution

Lane 1:

Human lymph node tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Lane 3:

Human lung tissue at 20 µg

Lane 4:

Human placenta tissue lysate at 20 µg

Lane 5:

Human colon tissue lysate at 20 µg

Lane 6:

Human testis tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 11 kDa,12 kDa,36 kDa

true

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-DAP12 antibody [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

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

In Western blot Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

All lanes:

Western blot - Anti-DAP12 antibody [EPR29151-41] (<a href='/en-us/products/primary-antibodies/dap12-antibody-epr29151-41-ab323318'>ab323318</a>) at 1/1000 dilution

Lane 1:

Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

THP-1 cells differentiated with TPA (12-O-Tetradecanoylphorbol-13-acetate) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 11 kDa,12 kDa,15 kDa

false

Exposure time: 26s

• Dot

Supplier Data

Dot Blot - Anti-DAP12 antibody [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

Dot blot analysis of DAP12 using ab323318 at 1000 (0.538 ug/ml) followed by a Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated (ab97051) at 1 : 100000 dilution.

Lane1 : Human DAP12 peptide
Lane2 : Human MTMR10 peptide

Exposure time : 180 seconds.

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

This antibody does not cross-react with human MTMR10.

All lanes:

Dot Blot - Anti-DAP12 antibody [EPR29151-41] (<a href='/en-us/products/primary-antibodies/dap12-antibody-epr29151-41-ab323318'>ab323318</a>) at 1/1000 dilution

Lane 1:

Human DAP12 peptide

Lane 2:

Human MTMR10 peptide

Secondary

All lanes:

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

false

Exposure time: 180s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DAP12 antibody [EPR29151-41] - BSA and Azide free (AB326592)

This data was developed using ab323318, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cells labelling DAP12 with ab323318 at 1/1000 (0.538 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing no staining in Jurkat cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Negative control : Jurkat (PMID : 9490415).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/ 200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

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