Anti-CD39 抗体 [EPR26473-58] (BSA and Azide free) (ab300066)

All lanes : Anti-CD39 antibody [EPR26473-58] (ab300065) at 1/1000 dilution

Lane 1 : IM-9 (Human multiple myeloma B Lymphoblast) whole cell lysate
Lane 2 : THP-1 (Human monocytic leukemia monocyte) whole cell lysate
Lane 3 : Jurkat ( Human T cell leukemia T lymphocyte) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 57 kDa
Observed band size: 78 kDa why is the actual band size different from the predicted?

This data was developed using ab300065, the same antibody clone in different buffer formulation. 

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

Lane 1 : Human placenta tissue lysate
Lane 2 : Human heart tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

Predicted band size: 57 kDa
Observed band size: 78 kDa why is the actual band size different from the predicted?

This data was developed using ab300065, the same antibody clone in different buffer formulation. 

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

Exposure time: 

Lane 1:59 seconds
Lane 2:180 seconds

This data was developed using ab300065, the same antibody clone in a different buffer formulation.

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

This data was developed using ab300065, the same antibody clone in a different buffer formulation.

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

This data was developed using ab300065, the same antibody clone in a different buffer formulation.

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

This data was developed using ab300065, the same antibody clone in a different buffer formulation.

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.

This data was developed using ab300065, the same antibody clone in a different buffer formulation.

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.

This data was developed using ab300065, the same antibody clone in a different buffer formulation.

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. 

This data was developed using ab300065, the same antibody clone in a different buffer formulation.

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

This data was developed using ab300065, the same antibody clone in a different buffer formulation.

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