Anti-CD39 antibody [EPR26473-58]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD39 antibody [EPR26473-58] (AB300065)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling CD39 with ab300065 at 1/2000 (0.295 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Positive staining on the endothelial cells and smooth muscle of human liver.The section was incubated with ab300065 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) was used.

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD39 antibody [EPR26473-58] (AB300065)

Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labelling CD39 with ab300065 at 1/2000 (0.295 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Positive staining on the stromal cells of human endometrial carcinoma (PMID : 25403716).The section was incubated with ab300065 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) was used.

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD39 antibody [EPR26473-58] (AB300065)

Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeilized IM-9 (human multiple myeloma B Lymphoblast) cells lebelling CD39 with ab300065 at 1/50 (11.8 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2μg/mL) dilution (Green). Confocal image showing membranous and cytoplasmic staining in IM-9 cell line. Negative control : Jurkat (PMID : 24319640). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5μg/ml) dilution (Red). 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 (2μg/mL) dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD39 antibody [EPR26473-58] (AB300065)

Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeilized Human PBMC cells lebelling CD39 with ab300065 at 1/50 (11.8 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2μg/mL) dilution (Green). Confocal image showing membranous and cytoplasmic staining in human PBMC is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 (2.5μg/ml) dilution (Red). 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 (2 μg/mL) dilution.

• Flow Cyt

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Flow Cytometry - Anti-CD39 antibody [EPR26473-58] (AB300065)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling CD39 with ab300065 at 1/500 dilution (0.1μg)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Cells were stained with rabbit IgG or ab300065. Then stained with anti-CD19 conjugated to Alexa Fluor^®647.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD39 antibody [EPR26473-58] (AB300065)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labelling with ab300065 at 1/2000 (0.295 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Positive staining on the endothelial cells and smooth muscle of human pancreas.The section was incubated with ab300065 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) was used.

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

• IP

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Immunoprecipitation - Anti-CD39 antibody [EPR26473-58] (AB300065)

CD39 was immunoprecipitated from 0.35 mg Human heart tissue lysate 10μg with ab300065 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300065 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Human heart tissue lysate 10μ

Lane 2 : ab300065 in Human heart tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300065 in Human heart tissue lysate

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

Exposure time : 67 seconds

All lanes:

Immunoprecipitation - Anti-CD39 antibody [EPR26473-58] (ab300065)

Predicted band size: 58 kDa

false

• IP

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Immunoprecipitation - Anti-CD39 antibody [EPR26473-58] (AB300065)

CD39 was immunoprecipitated from 0.35 mg Human placenta tissue lysate 10μg with ab300065 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300065 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Human placenta tissue lysate 10μg

Lane 2 : ab300065 in Human placenta tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300065 in Human placenta tissue lysate

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

Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-CD39 antibody [EPR26473-58] (ab300065)

Predicted band size: 58 kDa

false

• WB

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Western blot - Anti-CD39 antibody [EPR26473-58] (AB300065)

5% NFDM/TBST was used as blocking and diluting buffer.

Exposure time :

Lane 1 : 10 seconds
Lane 2 and 3 : 103 seconds

Negative control : Jurkat ( PMID : 9077545).

All lanes:

Western blot - Anti-CD39 antibody [EPR26473-58] (ab300065) at 1/1000 dilution

Lane 1:

IM-9 (Human multiple myeloma B Lymphoblast) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

Jurkat ( Human T cell leukemia T lymphocyte) 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 1/50000 dilution

Predicted band size: 58 kDa

Observed band size: 78 kDa

false

• WB

Supplier Data

Western blot - Anti-CD39 antibody [EPR26473-58] (AB300065)

5% NFDM/TBST was used as blocking and diluting buffer.

Exposure time :

Lane 1：59 seconds
Lane 2：180 seconds

All lanes:

Western blot - Anti-CD39 antibody [EPR26473-58] (ab300065) at 1/1000 dilution

Lane 1:

Human placenta tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 58 kDa

Observed band size: 78 kDa

false