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AB288330

Alexa Fluor® 647 Anti-CD28 antibody [EPR22076]

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Rabbit Recombinant Monoclonal CD28 antibody - conjugated to Alexa Fluor® 647. Suitable for ICC/IF, IHC-P and reacts with Human samples.

View Alternative Names

CD28, T-cell-specific surface glycoprotein CD28, TP44

2 Images
Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-CD28 antibody [EPR22076] (AB288330)
  • ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-CD28 antibody [EPR22076] (AB288330)

ab288330 staining CD28 in Jurkat cells, with negative expression in HEK293 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab288330 at 5 µg/ml (shown in red). ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488) (shown in green) was used at 2 µg/ml for structural counterstaining.

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-CD28 antibody [EPR22076] (AB288330)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-CD28 antibody [EPR22076] (AB288330)

Immunofluorescent staining of CD28 in a section of formalin-fixed paraffin-embedded human normal tonsil*.

Staining performed manually. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9.0) using retrieval settings of 110°C for 40 minutes on a Leica BOND. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab288330 at 1/100 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using DAKO Fluorescence Mounting Medium®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR22076

Isotype

IgG

Conjugation

Alexa Fluor® 647

Excitation/Emission

Ex: 650nm, Em: 665nm

Carrier free

No

Reacts with

Human

Applications

IHC-P, ICC/IF

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "5 µg/mL", "ICCIF-species-notes": "<p></p>", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/100", "IHCP-species-notes": "<p></p>" } } }
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We recommend this product because it’s often used in the same experiment or related research.

We advise that you always check the datasheet to ensure it fits your experiments, or contact ourtechnical teamfor help.

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Product details

This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.4 Preservative: 0.02% Sodium azide Constituents: 68% PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle|Store in the dark
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

CD28 also known as T-cell-specific surface glycoprotein CD28 or TP44 plays an important role as a co-stimulatory receptor on T cells. This glycoprotein has a molecular mass of about 44 kDa and is located mainly on the cell surface of T cells. CD28 expression also occurs on thymocytes and some B cells. It forms part of the immunoglobulin superfamily featuring in significant antibody interactions. Researchers often use anti-CD28 antibodies for various immunological assays including CD28 ELISA to study its function.
Biological function summary

CD28 co-stimulates T cells after antigen presentation. It enhances T cell proliferation survival and cytokine production. CD28 functions alongside the T cell receptor (TCR) complex mediating signals that promote full T cell activation. This interaction amplifies the immune response forming an important axis of immune system communication. CD28 interacts with its ligands B7-1 (CD80) and B7-2 (CD86) found on antigen-presenting cells (APCs) illustrating its role in immune system dynamics.

Pathways

CD28 is integral to the PI3K/Akt and MAPK/ERK signaling pathways. These pathways conduct signals critical for cellular growth and survival linking CD28 to downstream transcription factors. The interaction with another protein CTLA-4 acts as a regulatory balance for CD28 signaling preventing over-activation of T cells. This balance influences immune homeostasis and tolerance providing a framework for understanding costimulatory and inhibitory signaling.

CD28's role becomes evident in autoimmune diseases and transplant rejection. Abnormal CD28 signaling links to rheumatoid arthritis where inappropriate T cell activation occurs. Similarly CD28 activity contributes to organ transplant rejection by stimulating T cell-mediated responses against transplanted tissues. CTLA-4 through regulatory feedback can modify these effects providing therapeutic target strategies for managing such disorders. Efforts to modulate CD28 activity involve developing therapeutic agents such as anti-CD28 monoclonal antibodies.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Receptor that plays a role in T-cell activation, proliferation, survival and the maintenance of immune homeostasis (PubMed : 1650475, PubMed : 7568038). Functions not only as an amplifier of TCR signals but delivers unique signals that control intracellular biochemical events that alter the gene expression program of T-cells (PubMed : 24665965). Stimulation upon engagement of its cognate ligands CD80 or CD86 increases proliferation and expression of various cytokines in particular IL2 production in both CD4(+) and CD8(+) T-cell subsets (PubMed : 1650475, PubMed : 35397202). Mechanistically, ligation induces recruitment of protein kinase C-theta/PRKCQ and GRB2 leading to NF-kappa-B activation via both PI3K/Akt-dependent and -independent pathways (PubMed : 21964608, PubMed : 24665965, PubMed : 7568038). In conjunction with TCR/CD3 ligation and CD40L costimulation, enhances the production of IL4 and IL10 in T-cells (PubMed : 8617933).. Isoform 3. Enhances CD40L-mediated activation of NF-kappa-B and kinases MAPK8 and PAK2 in T-cells (PubMed : 15067037).
See full target information CD28
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions
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Associated Products

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Alternative Version
Primary Antibodies

AB243557

Anti-CD28 antibody [EPR22076] - BSA and Azide free

primary-antibodies

cd28-antibody-epr22076-bsa-and-azide-free-ab243557

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(1 reviews)

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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com