Anti-CD28 antibody [EPR22076]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD28 antibody [EPR22076] (AB243228)

Immunohistochemical analysis of paraffin-embedded human multiple myeloma tissue labeling CD28 with ab243228 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining on human Multiple Myeloma cells. (PMID : 7522634) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD28 antibody [EPR22076] (AB243228)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (human T cell leukemia T lymphocyte) cells labeling CD28 with ab243228 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in Jurkat cells. The nuclear counter stain is DAPI (blue).

Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).

The negative control is the secondary antibody only.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD28 antibody [EPR22076] (AB243228)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD28 with ab243228 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining on subsets of immune cells of human tonsil (PMID : 7489738) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IP

Supplier Data

Immunoprecipitation - Anti-CD28 antibody [EPR22076] (AB243228)

CD28 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte) whole cell lysate with ab243228 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab243228 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1 : Jurkat whole cell lysate 10 μg (Input).
Lane 2 : ab243228 IP in Jurkat whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab243228 in Jurkat whole cell lysate.
Blocking/Dilution buffer : 5% NFDM/TBST.
Exposure time : 30 seconds.

The target band ranging between 37kDa-48kDa is consistent with what has been described in the literature (PMID : 2825196). Minor band below 15kDa is likely to be an isoform.

All lanes:

Immunoprecipitation - Anti-CD28 antibody [EPR22076] (ab243228)

Predicted band size: 25 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD28 antibody [EPR22076] (AB243228)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD28 with ab243228 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining on subsets of immune cells of rat spleen is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD28 antibody [EPR22076] (AB243228)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD28 with ab243228 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining on subsets of immune cells of mouse spleen is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD28 antibody [EPR22076] (AB243228)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized EL4.IL-2 (Mouse lymphoma T lymphocyte) cells labeling CD28 with ab243228 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in EL4.IL-2 cells. The nuclear counter stain is DAPI (blue).

Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).

The negative control is the secondary antibody only.

• WB

Supplier Data

Western blot - Anti-CD28 antibody [EPR22076] (AB243228)

The target band ranging between 37kDa-48kDa is consistent with what has been described in the literature (PMID : 2825196). Minor band below 15kDa is likely to be an isoform.

This blot was developed using a higher sensitivity ECL substrate.

This primary antibody was incubated overnight at 4°C.

All lanes:

Western blot - Anti-CD28 antibody [EPR22076] (ab243228) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 2:

EL4.IL-2 (mouse lymphoma T lymphocyte), whole cell 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: 25 kDa

false

Exposure time: 26s