InnoSite™ iF488 Anti-human CD28 Antibody

Product Details


Clone028CB
ApplicationFlow Cytometry
ReactivityHuman
FormatiF488
Target NameCD28, Tp44
IsotypeRabbit IgG
Antibody TypeMonoclonal
Regulatory StatusRUO
FormulationPhosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 0.2% (w/v) BSA
Protein ConcentrationSupplied at a lot-specific concentration.
Storage & HandlingThe antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Recommended UsageFor flow cytometric staining, it is recommended to use 5 µL of this reagent per 0.5-1.0 million cells in a 100 µL volume. Optimal reagent performance should be determined by titration for each specific application. iF488 has an excitation max at 491 nm and an emission max at 516 nm.
Excitation LaserBlue Laser (488 nm)
BrandInnoSite
See All FormatsClone 028CB

Background Information


CD28 (Cluster of Differentiation 28) is a key costimulatory receptor expressed predominantly on T lymphocytes. It plays a vital role in the activation, proliferation, and survival of T-cells, complementing the antigen-specific signal delivered through the T-cell receptor (TCR). The engagement of CD28 is essential for full T-cell activation and the development of adaptive immune responses.

CD28 is a type I transmembrane glycoprotein belonging to the immunoglobulin (Ig) superfamily. Structurally, it exists as a homodimer on the T-cell surface and comprises three main parts: an extracellular Ig-like domain responsible for ligand binding, a transmembrane region, and a cytoplasmic tail containing signaling motifs. The intracellular region includes a YMNM motif that recruits PI3K (phosphoinositide 3-kinase) and adaptors such as Grb2 and GADS, promoting downstream signaling cascades including activation of Akt and NF-κB pathways. CD28 interacts primarily with B7 family ligands CD80 (B7-1) and CD86 (B7-2), expressed on professional antigen-presenting cells (APCs) such as dendritic cells, macrophages, and B cells. The binding of CD28 to these ligands, in conjunction with TCR engagement, enhances cytokine production (notably interleukin-2), cell metabolism, and survival, processes crucial for mounting effective immune responses.

Aberrant CD28 signaling is implicated in several pathological conditions. Overactivation can contribute to autoimmune diseases like rheumatoid arthritis and multiple sclerosis, where excessive T-cell stimulation leads to tissue damage. Conversely, reduced CD28 expression or signaling with aging (immunosenescence) can impair immune responses, leading to increased infection risk. Certain malignancies also exploit CD28 pathways to modulate immune evasion.

CD28 has been a therapeutic target and tool in immunotherapy. The development of CTLA-4-Ig fusion proteins (such as abatacept and belatacept) competitively blocks CD28-B7 interactions, reducing T-cell activation in autoimmune disorders and preventing transplant rejection. Conversely, engineered chimeric antigen receptor (CAR) T-cells often include CD28 intracellular domains to amplify antitumor activity by strengthening T-cell signaling. However, uncontrolled CD28 activation has proven hazardous, as exemplified by the TGN1412 trial, underscoring the need for careful modulation in therapeutic settings.

Isotype Control


InnoSite™ iF488 Rabbit IgG, κ2 Isotype Control

Data Sheets


InnoSite™ iF488 Anti-human CD28 Antibody TDS

Related Protocols


Flow Cytometry Protocol

Frequently Asked Questions


How do I determine the optimal antibody concentration/titer for my experiment?
We recommend titrating each new antibody lot on your own cell type and instrument, since optimal concentration depends on cell density, target expression level, and cytometer sensitivity. Suggested starting dilutions or test sizes (e.g., µg or µl per 10^6 cells) are provided on the product page as a starting reference point, not a fixed requirement.

Can these antibodies be used for intracellular or intranuclear staining, or only surface staining?
Intended use (surface, intracellular, or both) is specified per product, indicated under Application. The appropriate Related Protocol is linked for each product. If you need to detect both surface and intracellular markers in the same panel, confirm each antibody's compatibility with your fixation/permeabilization protocol before combining them.

What controls should I use alongside these antibodies?
Matched isotype controls (same host species, isotype, and conjugate) are available for most clones to help distinguish specific binding from background/non-specific staining and are linked on each product page. Unstained, single-color compensation, and fluorescence-minus-one (FMO) controls are also recommended, especially for multicolor panel design.

Can I combine InnoCyto Flow Cytometry antibodies into a multicolor panel?
Yes — antibodies across our catalog are designed to be panel-compatible, and conjugate options are offered specifically to support multiplexing. When building a panel, pair bright fluorophores with low-expression targets, avoid excessive spectral overlap, and confirm compensation is set up correctly for your specific fluorochrome combination. Use our Panel Builder to simplify the process.

Have a product or application question? Consult our FAQs or contact us.