Anti-PI 3 Kinase p110 delta antibody [EPR30972-532]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

Immunohistochemical analysis of paraffin-embedded Human Hodgkin lymphoma tissue labeling with ab327424 at 1/100 (5.19 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human Hodgkin lymphoma.

The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a human PIK3CD expression vector containing a Myc-His tag. (B) HEK-293T transfected with a human PIK3CB expression vector containing a Myc-His tag. (C) HEK-293T transfected with a human PIK3CA expression vector containing a Myc-His tag. (D) HEK-293T transfected with a human PIK3CG expression vector containing a Myc-His tag. (E) HEK-293T transfected with empty vector containing a Myc-His tag. tissue labeling with ab327424 at 1/2000 (0.26 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) HEK-293T transfected with a human PIK3CD expression vector containing a Myc-His tag, and no signal in (B) HEK-293T transfected with a human PIK3CB expression vector containing a Myc-His tag, (C) HEK-293T transfected with a human PIK3CA expression vector containing a Myc-His tag, (D) HEK-293T transfected with a human PIK3CG expression vector containing a Myc-His tag and (E) HEK-293T transfected with empty vector containing a Myc-His tag.

The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

Immunohistochemical analysis of paraffin-embedded Human thymoma tissue labeling with ab327424 at 1/100 (5.19 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human thymoma.

The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling with ab327424 at 1/100 (5.19 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human tonsil.

The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

Immunohistochemical analysis of paraffin-embedded (A) Raji (human Burkitt's lymphoma B lymphocyte) cell pellets (B) THP-1 (human monocytic leukemia monocyte) cell pellets (C) HEL (human erythroleukemia erythroblast) cell pellets (D) BxPC-3 (human pancreas adenocarcinoma epithelial cell) cell pellets tissue labeling with ab327424 at 1/500 (1.038 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in (A) Raji (human Burkitt's lymphoma B lymphocyte), (B) THP-1 (human monocytic leukemia monocyte)

and no staining in (C) HEL (human erythroleukemia erythroblast) and (D) BxPC-3 (human pancreas adenocarcinoma epithelial cell).

The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Raji (human Burkitt's lymphoma B lymphocyte, Right) BxPC-3 (human pancreas adenocarcinoma epithelial cell, Left) cells labelling with ab327424 at 1/50 dilution (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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : BxPC-3.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Right) NIH/3T3 (mouse embryonic fibroblast, Left) cells labelling with ab327424 at 1/50 dilution (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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : NIH/3T3.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling with ab327424 at 1/100 (5.19 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse spleen.

The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Lab

Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

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

Low expression : bEnd.3, CT26.WT, HEL, BxPC-3

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-3 : 180 seconds; Lanes 4-7 : 37 seconds

All lanes:

Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (ab327424) at 1/1000 dilution

Lane 1:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 48 µg

Lane 2:

bEnd.3 (mouse brain endothelial cell) whole cell lysate at 48 µg

Lane 3:

CT26.WT (mouse colon fibroblast) whole cell lysate at 48 µg

Lane 4:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 48 µg

Lane 5:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 48 µg

Lane 6:

HEL (human erythroleukemia erythroblast) whole cell lysate at 48 µg

Lane 7:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate at 48 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 119 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

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

Low expression : skeletal muscle, NIH/3T3

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-3 : 37 seconds; Lanes 4-5 : 180 seconds

All lanes:

Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (ab327424) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 48 µg

Lane 2:

Mouse thymus tissue lysate at 48 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 48 µg

Lane 4:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 48 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 48 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 119 kDa,36 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling with ab327424 at 1/100 (5.19 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in mouse skeletal muscle.

The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p110 delta antibody [EPR30972-532] (AB327424)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling with ab327424 at 1/100 (5.19 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in human skeletal muscle.

The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins