Rabbit anti-Mouse CD4 mAb - P06332

Core Product Specifications and Parameters

Biological Background: CD4 Function and Localization

• CD4, also known as L3T4 or Ly-4, is a T-cell surface glycoprotein essential for adaptive immunity. It serves as a co-receptor for MHC class II molecules on antigen-presenting cells.
• It interacts simultaneously with the T-cell receptor (TCR) and MHC class II:peptide complex, facilitating T-cell activation. Upon engagement, CD4 recruits the Src kinase LCK to the TCR-CD3 complex, initiating intracellular signaling cascades that lead to lymphokine production, adhesion, and T-helper cell activation.
• CD4 plays a critical role in thymic T-helper cell development and is also involved in the differentiation of monocytes into functional macrophages.
• Beyond T-cells, CD4 is expressed in other immune cells such as macrophages, dendritic cells, and NK cells, where it contributes to differentiation/activation, cytokine expression, and cell migration independently of TCR/LCK.
• The protein is a single-pass type I membrane glycoprotein with a molecular weight of approximately 51–55 kDa (UniProt: 51297 Da). It contains multiple immunoglobulin-like domains and is subject to post-translational modifications including N-glycosylation, phosphorylation, and palmitoylation.
• CD4 is highly expressed in T-helper cells, making it a defining marker for this lymphocyte subset. It is also present in regulatory T cells and a subset of innate lymphoid cells.
• Subcellular localization: primarily on the cell membrane. It cycles between the plasma membrane and endosomal compartments; palmitoylation aids in lipid raft localization, which is important for signaling.

Experimental Guidance and Technical Tips

• This antibody was raised against a recombinant protein corresponding to amino acids 27-394 of mouse CD4, covering the extracellular domain, and is validated for flow cytometry (FC). When designing experiments, consider titrating the antibody to optimize staining resolution, especially for rare cell populations.
• Since CD4 is expressed on T-helper cells and other immune cells, verify specificity in the appropriate mouse sample system (e.g., splenocytes, lymph node cells, or thymocytes). Use appropriate isotype controls and known CD4+ and CD4- cell lines for gating.
• For multicolor flow cytometry panels, ensure that the fluorophore conjugate is compatible with other markers of interest and that compensation controls are included.
• Although this product is validated for FC, its reactivity in other applications (e.g., IHC, WB) has not been tested; if required, users should perform in-house validation.

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