Invitrogen™ eBioscience™ CD28 Monoclonal Antibody (CD28.2), PE

The Invitrogen™ CD28 Monoclonal Antibody (CD28.2), conjugated to PE (Phycoerythrin), is a highly specific antibody designed for detecting human CD28 in flow cytometry applications. CD28, a 44 kDa homodimer, is a critical co-stimulatory molecule expressed on T cells, thymocytes, and plasma cells. It plays an essential role in T-cell activation by binding to CD80 (B7-1) and CD86 (B7-2) on antigen-presenting cells (APCs). This antibody is validated for intracellular and surface marker detection, providing bright fluorescence for reliable analysis.

Key Features:

1. Target and Specificity:
   • Recognizes human CD28, a key co-stimulatory molecule involved in T-cell activation, IL-2 production, and immune response regulation.
   • High specificity ensures accurate detection of CD28-expressing cells.
2. Clone and Conjugate:
   • Clone: CD28.2, a trusted and validated monoclonal antibody for CD28 detection.
   • Conjugate: PE (Phycoerythrin), providing bright fluorescence with excitation at 565 nm and emission at 576 nm.
3. Validated Applications:
   • Optimized for flow cytometry, enabling robust and reproducible detection of CD28 expression on T cells and plasma cells.
4. Bright and Stable Fluorescence:
   • PE ensures high signal intensity and minimal background, making it suitable for complex multicolor flow cytometry panels.
5. Ready-to-Use:
   • Pre-titrated and optimized for convenience, with a recommended usage of 5 µL (0.25 µg) per test.

Specifications:

Applications:

1. Flow Cytometry:
   • Detect surface expression of CD28 on T cells and plasma cells for phenotyping and functional studies.
2. T-cell Activation Studies:
   • Investigate CD28’s role in T-cell co-stimulation and its interaction with CD80/CD86 in immune responses.
3. Co-stimulatory Pathway Research:
   • Study the effects of CD28 signaling on IL-2 receptor expression, cytokine production, and T-cell proliferation.
4. Immune Regulation Research:
   • Examine CD28’s involvement in preventing T-cell anergy and promoting survival during immune responses.
5. Multicolor Flow Cytometry Panels:
   • Combine with other fluorochrome-conjugated antibodies for comprehensive immune cell profiling.

Key Benefits:

1. Bright and Consistent Fluorescence:
   • PE provides excellent brightness for clear and accurate identification of CD28-positive populations.
2. Validated and Reliable:
   • Tested for flow cytometry to ensure consistent and reproducible results across experiments.
3. Ease of Use:
   • Pre-titrated and ready-to-use formulation simplifies experimental workflows.
4. High Specificity:
   • Accurate detection of CD28 with minimal non-specific binding, even in complex staining protocols.
5. Compatible with Multicolor Panels:
   • PE’s spectral properties make it ideal for use alongside other fluorochrome-conjugated antibodies.

Usage Recommendations:

1. Sample Preparation:
   • Prepare peripheral blood mononuclear cells (PBMCs) or other immune cell samples for staining.
2. Staining Protocol:
   • Add 5 µL (0.25 µg) of antibody per 100 µL of cell suspension.
   • Incubate with cells in the dark at 4°C for 20–30 minutes.
3. Flow Cytometry:
   • Analyze using a flow cytometer equipped with a blue laser (488 nm) or yellow-green laser (561 nm).
4. Controls:
   • Include an appropriate isotype control (e.g., Mouse IgG1 kappa Isotype Control, PE) for gating and specificity validation.

Storage and Handling:

• Store at 4°C in the dark to protect the PE fluorochrome.
• Avoid freezing, as freezing may degrade the antibody or conjugate.

Target Information:

• CD28:
  • A 44 kDa disulfide-linked homodimer expressed on T cells, thymocytes, and plasma cells.
  • Binds to CD80 (B7-1) and CD86 (B7-2) on APCs, providing co-stimulatory signals required for T-cell activation.
  • Enhances IL-2 receptor expression, cytokine production, and T-cell proliferation.
  • Prevents T-cell anergy and supports immune regulation in diseases such as cancer and autoimmune disorders.

For Research Use Only:

This product is intended for research purposes only and is not for diagnostic or therapeutic applications.