Antibodies, Proteins, Kits and Reagents for Life Science

321 638 ₸ Product size

100 µl 321 638 ₸

Order now and get it on Tuesday March 02, 2021

Anti-CD28 antibody [EPR22076] (ab243228)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22076] to CD28
Suitable for: WB, IHC-P, ICC/IF, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-CD28 antibody [EPR22076]
See all CD28 primary antibodies
Description

Rabbit monoclonal [EPR22076] to CD28
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Mouse Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Jurkat and EL4.IL-2 whole cell lysate. IHC-P: Human multiple myeloma and tonsil tissue. Mouse and rat spleen tissue. ICC/IF: Jurkat and EL4.IL-2 cells. IP: Jurkat whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22076
Isotype

IgG
Research areas

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).
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).
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).
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).
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.
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.
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.
Western blot - Anti-CD28 antibody [EPR22076] (ab243228)

All lanes : Anti-CD28 antibody [EPR22076] (ab243228) at 1/1000 dilution

Lane 1 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lane 2 : EL4.IL-2 (mouse lymphoma T lymphocyte), whole cell 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: 25 kDa

Exposure time: 26 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.

This blot was developed using a higher sensitivity ECL substrate.

This primary antibody was incubated overnight at 4°C.
Anti-CD28 antibody [EPR22076] (ab243228)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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