Anti-CD28 antibody [EPR24592-5] - BSA and Azide free

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD28 antibody [EPR24592-5] - BSA and Azide free (AB283861)

This data was developed using ab283860, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of THP-1 (Human monocytic leukemia monocyte, left) / U266B1 (human multiple myeloma B lymphocyte, right) cells labelling CD28 with ab283860 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 Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : THP-1(PMID : 26773151)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD28 antibody [EPR24592-5] - BSA and Azide free (AB283861)

This data was developed using ab283860, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U266B1 cells labelling CD28 with ab283860 at 1/200 (2.675 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 membranous staining in U266B1 cells and no staining in 293T cells. Negative control : 293T(PMID : 27708164) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/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 2ug/ml dilution.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD28 antibody [EPR24592-5] - BSA and Azide free (AB283861)

This data was developed using ab283860, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling CD28 with ab283860 at 1/50 dilution (1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab283860. Then stained with anti-CD3 conjugated to Alexa Fluor® 647.Gated on viable cells.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD28 antibody [EPR24592-5] - BSA and Azide free (AB283861)

This data was developed using ab283860, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 293T (Human embryonic kidney epithelial cell, left) / Jurkat (Human T cell leukemia T lymphocyte, right) cells labelling CD28 with ab283860 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 Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : 293T(PMID : 27708164)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD28 antibody [EPR24592-5] - BSA and Azide free (AB283861)

This data was developed using ab283860, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat cells labelling CD28 with ab283860 at 1/200 (2.675 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 membranous staining in Jurkat cells and no staining in 293T cells. Negative control : 293T(PMID : 27708164) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/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 2ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-CD28 antibody [EPR24592-5] - BSA and Azide free (AB283861)

This data was developed using ab283860, the same antibody clone in a different buffer formulation.

CD28 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate 10 ug with ab283860 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283860 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate 10 ug

Lane 2 : ab283860 IP in Jurkat whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283860 in Jurkat whole cell lysate

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

Exposure time : 3 minutes

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Immunoprecipitation - Anti-CD28 antibody [EPR24592-5] (<a href='/en-us/products/primary-antibodies/cd28-antibody-epr24592-5-ab283860'>ab283860</a>)

Predicted band size: 25 kDa

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