CES3 Mouse mAb: Selection, Application & Protocol Guide
CES3 Mouse mAb: Selection, Application & Protocol Guide
CES3 Mouse mAb is a monoclonal antibody raised in mouse against carboxylesterase 3 (CES3), designed to support reliable immunodetection in a range of standard laboratory assay formats.
- Product SKU
- Ab350909
- Antibody Type
- Mouse Monoclonal Antibody (mAb)
- Formulation
- PBS only (no carrier protein, no azide)
- Target Protein
- CES3 (Carboxylesterase 3)
- Reference Catalog
- absea#KC-33880
- Recommended Storage
- Store at -20°C; avoid repeated freeze-thaw cycles
Introduction to CES3 and Its Biological Significance
Carboxylesterase 3 (CES3) belongs to the carboxylesterase (CES) gene family, a group of serine hydrolases responsible for the hydrolysis of ester and amide bonds in a broad range of endogenous lipids and xenobiotic compounds. CES enzymes are widely expressed in metabolically active tissues, including the liver, and play critical roles in lipid metabolism, drug detoxification, and prodrug activation.
CES3 has attracted growing research interest due to its potential involvement in triglyceride metabolism and its expression pattern in tissues relevant to metabolic disease. Accurate and reproducible detection of CES3 protein expression is therefore essential for studies in metabolic biology, hepatology, and pharmacology.
The CES3 Mouse mAb (Ab350909) provides a validated tool for specific detection of the CES3 protein, enabling researchers to interrogate expression levels, subcellular localization, and protein interactions in relevant experimental models.
Product Specifications and Formulation Considerations
The CES3 Mouse mAb (Ab350909, reference absea#KC-33880) is supplied as a monoclonal antibody of murine origin, formulated in phosphate-buffered saline (PBS) only. The absence of stabilizing carrier proteins (such as BSA or glycerol) and preservatives (such as sodium azide) in the formulation offers important practical advantages:
- Carrier-protein-free: Eliminates background interference in assays sensitive to exogenous proteins, such as ELISA and proximity ligation assays.
- Azide-free: Compatible with live-cell imaging, enzyme-linked detection systems, and HRP-conjugated secondary antibody systems, where azide would inhibit peroxidase activity.
- PBS-based buffer: Physiologically compatible and suitable for direct use or further dilution in standard assay buffers.
Researchers should aliquot the antibody upon first thaw to minimize freeze-thaw cycling and store working aliquots at 4°C for short-term use (up to two weeks). Long-term storage should be at -20°C or below.
Recommended Protocols and Application Guidelines
While specific validated application data should be confirmed with the datasheet supplied with Ab350909, CES3 Mouse mAbs of this class are generally suitable for use in the following applications. Researchers should optimize conditions empirically for their specific sample types and detection systems.
Western Blotting (WB)
Prepare whole-cell or tissue lysates under denaturing conditions using standard SDS-PAGE sample buffer. Recommended starting dilution: consult the product datasheet. Transfer to PVDF or nitrocellulose membrane, block with 5% non-fat dry milk or BSA in TBST, and incubate with primary antibody overnight at 4°C. Use an appropriate HRP-conjugated anti-mouse secondary antibody for detection.
Immunofluorescence (IF) / Immunocytochemistry (ICC)
Fix cells with 4% paraformaldehyde and permeabilize with 0.1–0.3% Triton X-100 if detecting intracellular targets. Block with 5% normal serum, then incubate with the primary antibody. Use a fluorophore-conjugated anti-mouse secondary antibody. Counterstain nuclei with DAPI as appropriate.
Immunoprecipitation (IP)
Pre-clear lysates with Protein A/G beads. Add the CES3 Mouse mAb at the manufacturer-recommended amount and incubate with rotation at 4°C. Capture immune complexes with Protein A/G beads, wash, and elute for downstream analysis.
Note: Optimal antibody concentrations for each application must be determined by the end user. Always include appropriate positive and negative controls.
Troubleshooting Common Issues
Even well-validated antibodies can yield suboptimal results if experimental conditions are not carefully controlled. The following troubleshooting guidance addresses the most frequent issues encountered when working with monoclonal antibodies such as CES3 Mouse mAb (Ab350909).
| Issue | Possible Cause | Recommended Action |
|---|---|---|
| No signal detected | Antibody concentration too low; inadequate antigen retrieval; target not expressed in sample | Increase antibody concentration; optimize antigen retrieval conditions; verify target expression with positive control lysate |
| High background | Insufficient blocking; antibody concentration too high; non-specific secondary antibody binding | Extend blocking time or increase blocker concentration; titrate primary antibody; use species-matched isotype control |
| Multiple non-specific bands (WB) | Sample degradation; cross-reactivity | Add protease inhibitors to lysis buffer; confirm sample integrity; review species cross-reactivity data on datasheet |
| Weak signal in IF | Over-fixation; poor permeabilization | Reduce fixation time; optimize detergent concentration for permeabilization |
Frequently asked questions
What species is the CES3 Mouse mAb (Ab350909) raised in, and what does this mean for secondary antibody selection?
The CES3 Mouse mAb (Ab350909) is a monoclonal antibody raised in mouse. For detection, you should use a secondary antibody directed against mouse immunoglobulins (e.g., anti-mouse IgG). Ensure the secondary antibody is compatible with your detection system (HRP, fluorophore, etc.) and that you use species-appropriate blocking reagents to minimize background.
Why is the CES3 Mouse mAb formulated in PBS only, and how does this affect my experimental workflow?
The PBS-only formulation means the antibody contains no carrier proteins (such as BSA) and no preservatives (such as sodium azide). This makes it ideal for enzyme-based detection systems where azide would inhibit HRP activity, and for assays where exogenous proteins could introduce background. However, because there are no stabilizers, it is important to aliquot the antibody immediately upon receipt and avoid repeated freeze-thaw cycles to maintain activity.
How should I determine the optimal working dilution for CES3 Mouse mAb in my specific assay?
Optimal working dilutions vary depending on the application (WB, IF, IP, etc.), the sample type, and the detection system used. Always start with the dilution range recommended on the product datasheet for Ab350909, then perform a titration experiment to identify the concentration that gives maximum specific signal with minimal background. Include a no-primary-antibody negative control in all titration experiments.
What positive control sample is recommended when using CES3 Mouse mAb?
A positive control should be a sample known to express CES3 at detectable levels. Liver-derived cell lines or tissue lysates are commonly used as positive controls for carboxylesterase family members due to their high expression in hepatic tissue. Consult the product datasheet for Ab350909 or published literature for confirmed positive control recommendations specific to this antibody.
References
- Aladdin Scientific. Ab350909 CES3 Mouse mAb product information page. (Internal reference: absea#KC-33880, PBS formulation).
- Holmes RS, Cox LA. Comparative structures and evolution of vertebrate carboxyl ester lipase (CEL) genes and proteins with a major role in reverse cholesterol transport. Cholesterol. 2012.
- Satoh T, Hosokawa M. Structure, function and regulation of carboxylesterases. Chem Biol Interact. 2006;162(3):195-211.
Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.