Recombinant 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
Related conjugates and formulations
Overview
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Product name
Anti-CD28 antibody [EPR22076]
See all CD28 primary antibodies -
Description
Rabbit monoclonal [EPR22076] to CD28 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IPmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22076 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry reagents
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab243228 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Predicted molecular weight: 25 kDa.
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IHC-P | (1) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/50.
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IP |
1/30.
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Notes |
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WB
1/1000. Predicted molecular weight: 25 kDa. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/50. |
IP
1/30. |
Target
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Function
Involved in T-cell activation, the induction of cell proliferation and cytokine production and promotion of T-cell survival. -
Tissue specificity
Expressed in T-cells and plasma cells, but not in less mature B-cells. -
Sequence similarities
Contains 1 Ig-like V-type (immunoglobulin-like) domain. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 940 Human
- Entrez Gene: 12487 Mouse
- Entrez Gene: 25660 Rat
- Omim: 186760 Human
- SwissProt: P10747 Human
- SwissProt: P31041 Mouse
- SwissProt: P31042 Rat
- Unigene: 591629 Human
see all -
Alternative names
- CD 28 antibody
- CD28 antibody
- CD28 antigen antibody
see all
Images
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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). -
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). -
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). -
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). -
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.
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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. -
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. -
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 secondsThe 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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (10)
ab243228 has been referenced in 10 publications.
- Luo Z et al. Ex vivo anchored PD-L1 functionally prevent in vivo renal allograft rejection. Bioeng Transl Med 7:e10316 (2022). PubMed: 36176616
- Ma YS et al. M2 macrophage-derived exosomal microRNA-155-5p promotes the immune escape of colon cancer by downregulating ZC3H12B. Mol Ther Oncolytics 20:484-498 (2021). PubMed: 33718596
- da Silva PHL et al. Presence of Senescent and Memory CD8+ Leukocytes as Immunocenescence Markers in Skin Lesions of Elderly Leprosy Patients. Front Immunol 12:647385 (2021). PubMed: 33777045
- Sajman J et al. Adhering interacting cells to two opposing coverslips allows super-resolution imaging of cell-cell interfaces. Commun Biol 4:439 (2021). PubMed: 33795833
- Perugino CA et al. CD4+ and CD8+ cytotoxic T lymphocytes may induce mesenchymal cell apoptosis in IgG4-related disease. J Allergy Clin Immunol 147:368-382 (2021). PubMed: 32485263
- Dong L et al. High Inflammatory Tendency Induced by Malignant Stimulation Through Imbalance of CD28 and CTLA-4/PD-1 Contributes to Dopamine Neuron Injury. J Inflamm Res 14:2471-2482 (2021). PubMed: 34140795
- Magiera-Mularz K et al. Human and mouse PD-L1: similar molecular structure, but different druggability profiles. iScience 24:101960 (2021). PubMed: 33437940
- Huang N et al. Splenic CD4+ and CD8+ T-cells highly expressed PD-1 and Tim-3 in cirrhotic patients with HCV infection and portal hypertension. Int J Immunopathol Pharmacol 35:20587384211061051 (2021). PubMed: 34930041
- Wang J & Xiong Y HSH2D contributes to methotrexate resistance in human T-cell acute lymphoblastic leukaemia. Oncol Rep 44:2121-2129 (2020). PubMed: 33000278
- Li Z et al. Exosomal miRNA-16-5p Derived From M1 Macrophages Enhances T Cell-Dependent Immune Response by Regulating PD-L1 in Gastric Cancer. Front Cell Dev Biol 8:572689 (2020). PubMed: 33330451