NFAT5 Mouse mAb: FAQs for Research Use

NFAT5 Mouse mAb is a monoclonal antibody raised in mouse and specifically designed to detect NFAT5, a tonicity-responsive transcription factor critical for cellular adaptation to hyperosmotic stress.

Product Name

NFAT5 Mouse mAb

SKU

Ab349695

Antibody Type

Monoclonal antibody, mouse-derived

Target Protein

NFAT5 (Nuclear Factor of Activated T-cells 5)

Biological Relevance

NFAT5 regulates gene expression in response to osmotic stress and is implicated in immune function, kidney physiology, and cancer biology

Supplier Reference

absea#KC-641

What Is NFAT5 and Why Is It Important?

NFAT5 (Nuclear Factor of Activated T-cells 5), also known as TonEBP (Tonicity-responsive Enhancer Binding Protein), is a transcription factor belonging to the Rel homology domain superfamily. Unlike other NFAT family members that are regulated by calcineurin-mediated dephosphorylation, NFAT5 is uniquely activated by hypertonic stress. Under hyperosmotic conditions, NFAT5 translocates to the nucleus and drives expression of genes encoding osmoprotective proteins, including those involved in organic osmolyte accumulation and heat-shock responses.

Beyond osmoregulation, NFAT5 plays roles in T-cell activation, macrophage function, tumor progression, and embryonic development. Its broad biological relevance makes it a valuable research target across nephrology, immunology, and oncology.

NFAT5 Mouse mAb: Key Product Information

The NFAT5 Mouse mAb (SKU: Ab349695) from Aladdin Scientific is a monoclonal antibody generated in mouse hosts and directed against NFAT5. It is supplied under the absea reference KC-641. Monoclonal antibodies offer high specificity and lot-to-lot consistency, making them preferred reagents for quantitative and reproducible assays.

For detailed information on host species reactivity, clone designation, immunogen sequence, isotype, conjugation state, and validated applications, please consult the product datasheet available on the Aladdin Scientific product page or contact our technical support team. Storage conditions, concentration, and volume details are provided on the product label and accompanying certificate of analysis.

Recommended Applications and Usage Guidelines

Monoclonal antibodies targeting NFAT5 are commonly employed in techniques such as Western blotting (WB), immunofluorescence (IF), immunohistochemistry (IHC), immunoprecipitation (IP), and flow cytometry (FC). The suitability of Ab349695 for each application should be confirmed against the validated applications listed on the official datasheet, as performance can vary by technique and sample type.

When designing experiments, researchers should include appropriate positive controls (e.g., cell lysates from hypertonic-stress-treated cells known to upregulate NFAT5) and negative controls (e.g., isotype control antibody or NFAT5-knockout cell lysate). Titration of primary antibody concentration is strongly recommended to optimize signal-to-noise ratio.

Storage, Handling, and Stability

Monoclonal antibodies are generally sensitive to repeated freeze-thaw cycles, which can lead to aggregation and loss of activity. Best practices include: aliquoting the antibody into single-use volumes upon first thaw, storing at the temperature specified on the datasheet (typically –20 °C for long-term or 4 °C for short-term use), and avoiding prolonged exposure to light if the antibody is fluorescently conjugated.

Do not use antibody preparations that appear turbid or precipitated without first clarifying by centrifugation, as aggregates may cause high background or inconsistent results. Always consult the product-specific datasheet for exact storage and stability recommendations for Ab349695.

Troubleshooting Common Issues

High background: Increase blocking stringency (e.g., use 5% BSA instead of non-fat milk for phospho-protein detection), optimize primary antibody dilution, or extend washing steps. Ensure secondary antibody does not cross-react with the sample species.

No signal or weak signal: Verify that NFAT5 is expressed in your cell or tissue model under the chosen conditions—consider inducing expression with hypertonic stress (e.g., NaCl or sorbitol treatment). Check antibody lot viability, confirm correct secondary antibody pairing, and ensure sufficient antigen retrieval for IHC applications.

Non-specific bands (Western blot): Use a validated positive control lysate, reduce primary antibody concentration, and confirm sample preparation did not degrade the target. Consulting the Aladdin Scientific technical support team is recommended when standard troubleshooting steps do not resolve the issue.

Frequently asked questions

What applications is the NFAT5 Mouse mAb (Ab349695) validated for?

The specific validated applications for Ab349695 are detailed in the product datasheet. Commonly, NFAT5 monoclonal antibodies are used in Western blotting, immunofluorescence, immunohistochemistry, immunoprecipitation, and flow cytometry. Please consult the official Aladdin Scientific product page or contact technical support for a confirmed application list.

Which species does the NFAT5 Mouse mAb (Ab349695) recognize?

Species reactivity for Ab349695 is specified in the product datasheet. Researchers should verify that the antibody has been validated for the organism of interest prior to use, as cross-reactivity is not guaranteed across all species. Contact Aladdin Scientific technical support if you need guidance on reactivity with a specific model organism.

How should I store the NFAT5 Mouse mAb to maintain activity?

Monoclonal antibodies should generally be stored at –20 °C for long-term use and at 4 °C for short-term use, with repeated freeze-thaw cycles avoided by preparing single-use aliquots. Always refer to the specific storage instructions on the Ab349695 product label and datasheet, as the manufacturer's guidance takes precedence.

What positive control should I use when testing the NFAT5 Mouse mAb?

A suitable positive control is a lysate from cells known to express NFAT5, particularly those subjected to hypertonic stress conditions (such as treatment with high NaCl or sorbitol concentrations), which upregulate NFAT5 expression. NFAT5-overexpressing cell lines or tissues with known high NFAT5 expression, such as renal medulla, may also serve as positive controls.

Can I use the NFAT5 Mouse mAb for immunoprecipitation experiments?

Whether Ab349695 is validated for immunoprecipitation should be confirmed on the product datasheet. If the application is listed as validated, standard IP protocols optimized for monoclonal antibodies can be followed. If IP is not listed, contact Aladdin Scientific technical support to discuss feasibility or alternative product options.

What is the difference between NFAT5 and other NFAT family members?

NFAT5 is unique within the NFAT family because its activation is driven primarily by hyperosmotic stress rather than by calcium-calcineurin signaling, which governs NFAT1–4. NFAT5 also lacks the regulatory serine-rich domains targeted by calcineurin and is constitutively localized in a partially nuclear fashion, further distinguishing it from classical NFAT members.

References

1. Burg MB, Ferraris JD, Dmitrieva NI. Cellular response to hyperosmotic stresses. Physiological Reviews. 2007;87(4):1441-1474.
2. Lopez-Rodriguez C, Aramburu J, Jin L, Bhatt A, Darieva Z, Bhatt DL, Rao A. Bridging the NFAT and NF-κB families: NFAT5 dimerization regulates cytokine expression in T cells. Nature Immunology. 2001;2(2):149-156.
3. Yancey PH. Organic osmolytes as compatible, metabolic and counteracting cytoprotectants in high osmolarity and other stresses. Journal of Experimental Biology. 2005;208(Pt 15):2819-2830.

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