Anti-Human Fibulin-4 [Clone 5G11] — Purified in vivo GOLD™ Functional Grade

Clone 5G11b is primarily used in in vivo mouse studies to target the lymphotoxin beta receptor (LT?R) for functional activation, allowing investigation of LT?R-associated signaling and immunological responses.

This monoclonal antibody, specifically rat anti-mouse LT?R (clone 5G11b), is agonistic: it activates LT?R signaling in vivo, particularly via the NF-?B pathway, and induces downstream effects such as MIP-2 chemokine induction and IL-6 release. These features make it suitable for studying roles of LT?R in immune regulation, stromal cell activation, and inflammation within live mouse models. The antibody is frequently used in experiments where researchers need to stimulate LT?R signaling (as opposed to blocking it), allowing assessment of receptor function in physiological or disease contexts such as autoimmune disease, lymphoid organ development, or response to infection.

Key points about its use:

• Functional activation (agonism): The antibody drives LT?R-linked signaling, unlike blocking antibodies.
• Cellular targets: LT?R is primarily expressed on stromal cells and fibroblasts, and at lower levels on certain myeloid cell lines.
• Assays and endpoints: Studies typically measure NF-?B activation, chemokine/cytokine induction, and functional outcomes (such as immune cell recruitment or tissue changes).
• The antibody is routinely tested for activity on WEH164S mouse cells, ensuring consistent agonistic properties.

If you were asking about desmoglein 3 clone 5G11 (instead of LT?R 5G11b), note that this is used mainly for immunostaining human tissues and does not pertain to in vivo mouse activation studies.

In summary, clone 5G11b is used in mouse studies as a functional agonist to stimulate lymphotoxin beta receptor activity, facilitating studies on immune signaling and tissue responses in vivo.

The correct storage temperature for a sterile packaged clone 5G11 depends on the specific antibody product in question, as "clone 5G11" refers to a particular hybridoma line and can be used for several different monoclonal antibodies.

• Anti-TOPORS mouse monoclonal antibody (clone 5G11): Should be stored at -20°C or -80°C in PBS (pH 7.4), with repeated freeze-thaw cycles to be avoided.
• Anti-Desmoglein-3 (clone 5G11): Stable for up to one year at 2–8°C from the date of receipt.
• Anti-Streptococcus phocae (clone 5G11/B6): Should be stored at -20°C and is stable for at least one year at this temperature, with freeze-thaw cycles to be avoided.

Conclusion:
There is no universal storage temperature for all sterile packaged clones labeled 5G11—you must refer to the specific product’s documentation. If you are unsure which product you have, consult the supplier or product datasheet to confirm the correct storage conditions for your particular monoclonal antibody. Most monoclonal antibodies are stored at 2–8°C for short-term use or at -20°C for longer storage, with aliquoting and avoidance of freeze-thaw cycles as general best practices. Always check the manufacturer’s instructions for your specific clone.

In the context of antibodies and proteins used alongside the 5G11 clone, the literature primarily focuses on the specific targets each antibody recognizes rather than direct pairings. However, it is common for researchers to use multiple antibodies or proteins in studies to achieve comprehensive understanding or to target different aspects of a biological system. Here are some examples of commonly used antibodies or proteins in research:

1. Desmoglein-3 Antibody (5G11 Clone)

   • Target: Desmoglein-3
   • Use: Immunofluorescence, Immunohistology, Western Blotting
   • Specificity: Recognizes human desmoglein-3, a component of desmosomes in epithelial cells.

2. Fibulin-4 Antibody (5G11 Clone)

   • Target: Fibulin-4
   • Use: Immunofluorescence, Immunohistochemistry, Western Blotting
   • Specificity: Recognizes human Fibulin-4, involved in extracellular matrix interactions.

3. Other Antibodies and Proteins

   • Monobodies, Pronectins, Adnectins: These are non-antibody proteins used for targeted binding, similar to antibodies but derived from different scaffolds.
   • Affibodies: Derived from protein A, used for targeted binding in therapeutic applications.
   • Anticalins: Derived from lipocalin proteins, used for targeted binding with high affinity and specificity.
   • Cytokine Muteins: Engineered cytokines with reduced binding affinity, used in combination with targeting antibodies for therapeutic applications.

In practice, researchers may use combinations of monoclonal antibodies and other targeted proteins to achieve specificity in binding to different epitopes or targets within a biological system. However, the specific pairing of these with the 5G11 clone is not directly detailed in the provided literature but would depend on the research goals and the specific antigens being targeted.

Key Findings from Clone 5G11 (Anti-Desmoglein 3 Antibody) in Scientific Literature

Specificity Epitope

• Clone 5G11 is described as a monoclonal mouse anti-human desmoglein 3 (DSG3) antibody that specifically targets the terminal extracellular (EC) portion of human DSG3, specifically amino acids 446–613.
• This is relevant because DSG3 is a key autoantigen in pemphigus vulgaris (PV), an autoimmune blistering disease. Most pathogenic anti-DSG3 antibodies bind to the N-terminal EC1–EC2 domains, but 5G11 binds a different region.
• By contrast, mapping studies with 5G11 have not identified a specific linear epitope; its binding is to a conformational, terminal EC region, distinct from the EC1–EC2 domains targeted by PV-causing antibodies.

Non-pathogenic Binding

• 5G11 was used as a primary antibody in immunohistochemistry/immunofluorescence, confirming its specificity for human DSG3 in tissue sections and cell-based assays.
• Experimental context: 5G11 was employed as a negative control in epitope mapping for pathogenic antibodies, highlighting its non-pathogenic nature—it did not recognize linear synthetic peptides associated with the pathogenic epitopes.
• Unlike pathogenic clones (e.g., AK23), 5G11 does not cause keratinocyte dissociation (a hallmark of PV pathogenicity), further confirming it lacks pathogenic potential in cellular assays.

Species Specificity

• The literature notes that 5G11 does not cross-react with mouse DSG3. This is relevant for experimental design, as some studies generate humanized or transgenic mouse models expressing human DSG3, where 5G11 is used to specifically detect the human protein.
• In transgenic models (expressing human DSG3 in mice), 5G11 is used to confirm expression of the human transgene and to distinguish it from endogenous mouse DSG3.

Summary Table

Conclusion

Clone 5G11 is a well-characterized, non-pathogenic, human-specific monoclonal antibody targeting the terminal extracellular domain of DSG3. It is widely used in research settings to specifically detect human DSG3 expression, especially in transgenic models, and serves as a negative control in studies distinguishing pathogenic from non-pathogenic anti-DSG3 antibodies. Its lack of cross-reactivity with mouse DSG3 and inability to induce PV-like pathology make it a valuable tool for DSG3 research.