IL12A Mouse mAb Protocol: Applications & Usage Guide

IL12A Mouse mAb is a monoclonal antibody targeting the IL-12 p35 subunit (IL12A), supplied in PBS buffer and suitable for immunological research applications requiring high specificity against human or relevant species IL-12A antigen.

Product SKU

Ab350096

Antibody Type

Mouse Monoclonal Antibody (mAb)

Target Antigen

IL12A (IL-12 p35 subunit)

Formulation

PBS (Phosphate-Buffered Saline) only

Host Species

Mouse

Reference Clone

absea#KC-469

Background: IL-12A and Its Biological Significance

Interleukin-12 subunit alpha (IL12A), also known as the p35 subunit, is one of the two subunits that form the biologically active heterodimeric cytokine IL-12 (p70). IL-12 is primarily produced by dendritic cells, macrophages, and B cells in response to antigenic stimulation and plays a central role in bridging innate and adaptive immunity. IL-12 drives the differentiation of naïve CD4+ T helper cells toward the Th1 phenotype and promotes the cytotoxic activity of CD8+ T lymphocytes and NK cells, making it a key target in cancer immunology, infectious disease research, and autoimmune studies.

The IL12A Mouse mAb (Ab350096) is designed to specifically recognize the IL-12A p35 subunit, providing a reliable tool for detecting, quantifying, or neutralizing IL-12A in experimental settings. Its monoclonal nature ensures lot-to-lot consistency and high epitope specificity.

Recommended Protocol & Working Concentrations

The following general protocol guidelines apply to the IL12A Mouse mAb (Ab350096). Always perform application-specific optimization in your laboratory, as optimal conditions may vary depending on sample type, instrument, and detection system.

General Preparation

• Briefly centrifuge the vial before opening to collect contents at the bottom.
• Reconstitute or dilute as required using PBS (pH 7.2–7.4) or an appropriate assay buffer. No carrier protein is required if the formulation is PBS-only.
• Prepare working dilutions fresh on the day of use. Avoid repeated freeze-thaw cycles.

Suggested Application Dilutions (to be validated by user)

• ELISA: Titrate starting from 1–2 µg/mL; optimize capture and detection concentrations empirically.
• Western Blot (WB): Start at 0.5–2 µg/mL in blocking buffer (5% non-fat dry milk or BSA in TBST).
• Immunofluorescence (IF) / Immunohistochemistry (IHC): Start at 2–5 µg/mL; antigen retrieval may be required for fixed tissues.
• Flow Cytometry: Titrate 0.5–2 µg per 10⁶ cells; use appropriate secondary antibody conjugate.

Note: Specific validated dilutions are not confirmed in the current source material. Users should perform a dilution series to establish optimal working concentration for each application.

Storage, Stability & Handling

Proper storage and handling of the IL12A Mouse mAb are essential to maintain antibody activity and ensure reproducible results.

• Short-term storage (≤1 month): Store at 4°C. Keep the vial tightly sealed and protected from light.
• Long-term storage: Store at −20°C or −80°C in single-use aliquots to avoid repeated freeze-thaw cycles, which can lead to antibody aggregation and loss of activity.
• Formulation: PBS only (no preservative or carrier protein specified in source data). Consider adding a carrier protein such as 0.1% BSA if long-term stability at 4°C is required.
• Do not use if the solution appears turbid or if the vial has been improperly stored.
• Avoid azide-containing buffers if the antibody is intended for cell-based functional assays, as azide is toxic to cells.

Note: Specific expiry data and stability study results are not provided in the current source material. Refer to the product datasheet (Ab350096) for lot-specific information.

Troubleshooting Common Issues

The following troubleshooting guide addresses common challenges encountered when using monoclonal antibodies such as IL12A Mouse mAb (Ab350096):

Problem	Possible Cause	Recommended Action
No signal detected	Antibody concentration too low; incompatible secondary antibody; antigen not expressed in sample	Increase primary antibody concentration; verify secondary antibody species compatibility (anti-mouse); confirm IL12A expression in your cell/tissue model
High background	Insufficient blocking; primary antibody concentration too high; non-specific binding	Optimize blocking (5% BSA or milk); reduce primary antibody concentration; increase wash stringency
Weak or inconsistent signal	Repeated freeze-thaw degradation; suboptimal antigen retrieval	Use freshly thawed aliquots; optimize antigen retrieval method for IHC/IF applications
Unexpected band size (WB)	Post-translational modifications; protein complexes; non-specific binding	Verify expected molecular weight of IL12A (~35 kDa for p35 subunit); include positive control lysate

Frequently asked questions

What is the formulation of IL12A Mouse mAb (Ab350096)?

The IL12A Mouse mAb (Ab350096) is formulated in PBS (Phosphate-Buffered Saline) only, with no carrier protein or preservative specified in the current product information. Researchers requiring long-term 4°C storage may consider supplementing with 0.1% BSA.

Which applications is the IL12A Mouse mAb suitable for?

While specific validated applications are not confirmed in the current source material, mouse monoclonal antibodies targeting IL12A are commonly used in ELISA, Western blot, immunofluorescence, immunohistochemistry, and flow cytometry. Users should validate the antibody for their specific application and sample type. Please refer to the product datasheet for Ab350096 for confirmed application data.

How should I store the IL12A Mouse mAb to maintain its activity?

For short-term use (up to one month), store the antibody at 4°C in a sealed, light-protected vial. For long-term storage, aliquot the antibody into single-use volumes and store at −20°C or −80°C to prevent activity loss from repeated freeze-thaw cycles. Always consult the product datasheet for lot-specific recommendations.

What is the clone reference for IL12A Mouse mAb (Ab350096)?

The IL12A Mouse mAb (Ab350096) is referenced as clone absea#KC-469. This clone designation can be used when citing the reagent in publications or when contacting technical support for clone-specific information.

How do I troubleshoot high background signal when using IL12A Mouse mAb?

High background is typically caused by insufficient blocking, excessively high primary antibody concentration, or inadequate washing. To resolve this, optimize your blocking step using 5% BSA or non-fat dry milk, reduce the primary antibody concentration, and increase the number or duration of wash steps. Ensure that the secondary antibody is specific to mouse IgG to minimize cross-reactivity.

References

1. Trinchieri G. Interleukin-12 and the regulation of innate resistance and adaptive immunity. Nature Reviews Immunology. 2003;3(2):133–146. doi:10.1038/nri1001
2. Kobayashi M, et al. Identification and purification of natural killer cell stimulatory factor (NKSF), a cytokine with multiple biologic effects on human lymphocytes. Journal of Experimental Medicine. 1989;170(3):827–845.
3. Aladdin Scientific. IL12A Mouse mAb (Ab350096) Product Datasheet. Aladdin Scientific.

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.