Anti-CD73 antibody [EPR25191-104] - BSA and Azide free

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD73 antibody [EPR25191-104] - BSA and Azide free (AB288161)

This data was developed using ab288154, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of Mouse splenocytes cells labelling CD73 with ab288154 at 1/500 dilution (0.1ug) (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.

Cells were stained with rat IgG or ab288154. Then stained with anti-CD3 conjugated to Alexa Fluor^® 647. Gated on viable cells.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD73 antibody [EPR25191-104] - BSA and Azide free (AB288161)

This data was developed using ab288154, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage, Left) / 4T1 (Mouse mammary gland carcinoma epithelial cell, Right) cells labelling CD73 with ab288154 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). Goat Anti-Rabbit IgG (Alexa Fluo 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

Negative control : RAW 264.7(PMID : 28060378).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD73 antibody [EPR25191-104] - BSA and Azide free (AB288161)

This data was developed using ab288154, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 4T1 cells labelling CD73 with ab288154 at 1/500 (1.038 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in 4T1 cells and no staining in RAW 264.7 cells. Negative control : RAW264.7(PMID : 28060378). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 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 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-CD73 antibody [EPR25191-104] - BSA and Azide free (AB288161)

This data was developed using ab288154, the same antibody clone in a different buffer formulation.

CD73 was immunoprecipitated from 0.35 mg 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate with ab288154 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288154 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab288154 IP in 4T1 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288154 in 4T1 whole cell lysate

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

Exposure time : 41 seconds

All lanes:

Immunoprecipitation - Anti-CD73 antibody [EPR25191-104] (<a href='/en-us/products/primary-antibodies/cd73-antibody-epr25191-104-ab288154'>ab288154</a>)

Predicted band size: 63 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-CD73 antibody [EPR25191-104] - BSA and Azide free (AB288161)

This data was developed using ab288154, the same antibody clone in a different buffer formulation.

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

Exposure time : 81 seconds

All lanes:

Western blot - Anti-CD73 antibody [EPR25191-104] (<a href='/en-us/products/primary-antibodies/cd73-antibody-epr25191-104-ab288154'>ab288154</a>) at 1/1000 dilution

Lane 1:

Mouse bladder tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 63 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-CD73 antibody [EPR25191-104] - BSA and Azide free (AB288161)

This data was developed using ab288154, the same antibody clone in a different buffer formulation.

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

The observed MW/expression profile is consistent with what has been described in the literature (PMID : 28399915;21533188).

Negative control : RAW264.7, J774A.1 (PMID : 28060378)

Exposure time : 81 seconds

All lanes:

Western blot - Anti-CD73 antibody [EPR25191-104] (<a href='/en-us/products/primary-antibodies/cd73-antibody-epr25191-104-ab288154'>ab288154</a>) at 1/1000 dilution

Lane 1:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

J774A.1 (mouse reticum cell sarcoma monocyte macrophage) 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

Predicted band size: 63 kDa

Observed band size: 70 kDa

false