S100 Alpha6 Mouse mAb: Selection & Application Guide

S100 alpha6 Mouse mAb is a mouse-derived monoclonal antibody specifically targeting S100A6, a member of the S100 calcium-binding protein family, formulated in PBS and supplied by Aladdin Scientific as catalog Ab351066.

Catalog Number

Ab351066

Antibody Type

Mouse Monoclonal Antibody (mAb)

Target Antigen

S100 alpha6 (S100A6)

Formulation

PBS (Phosphate-Buffered Saline) only

Supplier Reference

absea#KC-46664

Species Reactivity

To be confirmed; please refer to datasheet

Overview of S100 Alpha6 and Its Biological Significance

S100 alpha6, also known as S100A6 or calcyclin, is a low-molecular-weight calcium-binding protein belonging to the S100 protein family. S100 proteins are characterized by two EF-hand calcium-binding domains and are involved in the regulation of a wide range of intracellular processes, including cell cycle progression, differentiation, apoptosis, and cytoskeletal dynamics.

S100A6 is expressed in a variety of cell types and is notably upregulated in several human cancers, making it a subject of interest in oncology research. It has also been studied in the context of neurological conditions and inflammatory responses. Its dual roles — both intracellular signaling and extracellular secretion — make it a versatile and important research target.

Using a high-quality, well-characterized monoclonal antibody such as Ab351066 is essential for obtaining reproducible and specific results when studying S100A6 expression and function.

Product Specifications and Formulation Details

Ab351066, the S100 alpha6 Mouse Monoclonal Antibody from Aladdin Scientific, is produced using standard hybridoma technology and is formulated in PBS (phosphate-buffered saline) without additional stabilizers, preservatives, or carrier proteins unless otherwise stated on the datasheet. The PBS-only formulation offers flexibility for downstream conjugation, labeling, or custom formulation by the end user.

Key specifications to confirm on the datasheet include:

• Immunogen used for immunization
• Isotype (e.g., IgG subclass)
• Concentration and total volume per vial
• Validated application types (e.g., WB, IHC, IF, ELISA, Flow Cytometry)
• Species reactivity (e.g., human, mouse, rat)
• Storage conditions and recommended working dilutions

Please consult the official product datasheet for Ab351066 for the most current and complete specifications, as these details are subject to lot-to-lot confirmation.

Recommended Applications and Usage Protocol

Mouse monoclonal antibodies targeting S100A6 are commonly used in a range of immunological and biochemical detection techniques. While the specific validated applications for Ab351066 should be confirmed on the datasheet, the following general protocols apply to S100A6 mouse mAbs in research settings:

Western Blotting (WB)

Denature samples under reducing conditions using SDS-PAGE. Transfer to PVDF or nitrocellulose membrane. Block with 5% non-fat dry milk or BSA in TBST for 1 hour at room temperature. Incubate with primary antibody at recommended dilution (typically 1:500–1:2000) overnight at 4°C. Wash and apply appropriate HRP-conjugated secondary anti-mouse antibody. Detect by chemiluminescence.

Immunohistochemistry (IHC)

Perform antigen retrieval on FFPE sections (citrate buffer pH 6.0 or EDTA buffer pH 9.0 is commonly used for S100 family proteins). Block endogenous peroxidase activity and non-specific binding. Apply primary antibody at appropriate dilution and incubate. Use a polymer-based detection system with DAB chromogen and hematoxylin counterstain.

Immunofluorescence (IF)

Fix cells with 4% paraformaldehyde and permeabilize with 0.1–0.3% Triton X-100. Block with 5% normal serum. Apply primary antibody and incubate at 4°C overnight. Detect with a fluorophore-conjugated anti-mouse secondary antibody and counterstain nuclei with DAPI.

Note: Always titrate the antibody for each specific application and cell/tissue type to determine optimal working concentration.

Storage, Handling, and Stability

Proper storage and handling of Ab351066 are critical for maintaining antibody activity and ensuring reproducible experimental results. As the product is formulated in PBS only, the following general guidelines apply:

• Short-term storage (up to 1 month): Store at 4°C. Avoid repeated freeze-thaw cycles.
• Long-term storage: Aliquot the antibody into single-use volumes and store at -20°C or -80°C as indicated on the datasheet. Addition of a cryoprotectant such as glycerol (to 50% v/v) may be advisable for long-term storage at -20°C if not already included in the formulation.
• Avoid: Exposure to repeated freeze-thaw cycles, prolonged exposure to light (if fluorescently conjugated), and contamination with proteases or detergents.
• Handling: Centrifuge briefly before opening to ensure the product is at the bottom of the vial. Work on ice during experiment setup.

Always refer to the lot-specific datasheet supplied with Ab351066 for definitive storage instructions, expiry date, and concentration information.

Troubleshooting Common Issues

Researchers using Ab351066 may occasionally encounter suboptimal results. The following troubleshooting guide addresses the most common issues:

Problem	Possible Cause	Suggested Solution
No or weak signal	Antibody concentration too low; antigen retrieval insufficient; target not expressed in sample	Increase antibody concentration; optimize antigen retrieval conditions; use positive control tissue or cell line known to express S100A6
High background	Insufficient blocking; antibody concentration too high; non-specific secondary antibody binding	Extend blocking time or change blocking agent; reduce primary antibody concentration; ensure secondary antibody is species-matched
Non-specific bands (WB)	Sample overloading; cross-reactivity with related S100 family members	Reduce protein loading; include a negative control lysate; verify band size against expected molecular weight of S100A6 (~10–11 kDa)
Loss of signal after storage	Repeated freeze-thaw cycles; improper storage temperature	Aliquot antibody upon receipt; store at recommended temperature; avoid freeze-thaw cycles

Contact Aladdin Scientific technical support if issues persist after troubleshooting steps have been applied.

Frequently asked questions

What is S100 alpha6 (S100A6) and why is it an important research target?

S100A6 is a member of the S100 family of EF-hand calcium-binding proteins involved in regulating cell cycle, differentiation, and apoptosis. It is of significant research interest because it is upregulated in multiple cancer types and has roles in neurological and inflammatory pathways, making it a potential biomarker and therapeutic target.

What formulation is Ab351066 supplied in, and does it contain preservatives?

Ab351066 is supplied in PBS (phosphate-buffered saline) only. Based on the available product information, no additional preservatives, stabilizers, or carrier proteins are indicated. Users requiring a specific formulation should contact Aladdin Scientific for further details or consider custom dialysis.

Which experimental applications is Ab351066 suitable for?

S100A6 mouse monoclonal antibodies are commonly used in Western blotting (WB), immunohistochemistry (IHC), immunofluorescence (IF), and ELISA. The specific validated applications for Ab351066 are detailed in the product datasheet; researchers should consult the datasheet to confirm suitability for their intended application.

How should Ab351066 be stored to maintain optimal activity?

For short-term use (up to one month), store at 4°C. For long-term storage, aliquot into single-use volumes and store at -20°C or -80°C to avoid repeated freeze-thaw cycles, which can degrade antibody performance. Always refer to the lot-specific datasheet for definitive storage recommendations.

How do I select the appropriate working dilution for Ab351066?

Working dilutions vary depending on the application (WB, IHC, IF, etc.) and the specific sample type. The recommended starting dilutions are provided on the product datasheet. It is always advisable to perform an initial titration experiment using positive control samples to determine the optimal dilution for your experimental conditions.

References

1. Donato R. S100: a multigenic family of calcium-modulated proteins of the EF-hand type with intracellular and extracellular functional roles. International Journal of Biochemistry & Cell Biology. 2001;33(7):637-668. doi:10.1016/S1357-2725(01)00046-2
2. Brozzi F, Arcuri C, Giambanco I, Donato R. S100B protein regulates astrocyte shape and migration via interaction with Src kinase: implications for astrocyte development, activation, and tumor growth. Journal of Biological Chemistry. 2009;284(13):8797-8811.
3. Salama I, Malone PS, Mihaimeed F, Jones JL. A review of the S100 proteins in cancer. European Journal of Surgical Oncology. 2008;34(4):357-364. doi:10.1016/j.ejso.2007.04.009

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