SNX25 Mouse mAb: Selection, Application & Protocol Guide

SNX25 Mouse mAb is a mouse-derived monoclonal antibody targeting the sorting nexin 25 (SNX25) protein, designed to support researchers in investigating endosomal sorting, membrane trafficking, and related cellular processes with high specificity and reproducibility.

Product SKU

Ab351394

Antibody Type

Mouse Monoclonal Antibody (mAb)

Formulation

PBS only (no carrier protein, no preservative)

Target Protein

SNX25 (Sorting Nexin 25)

Host Species

Mouse

Reference Catalog

absea #KC-31961

Overview of SNX25 and Antibody Rationale

Sorting nexin 25 (SNX25) is a member of the sorting nexin family, a group of proteins characterized by the presence of a phox homology (PX) domain that mediates binding to phosphoinositide lipids on endosomal membranes. SNX25 has been implicated in endosomal sorting pathways and membrane trafficking, playing roles in the regulation of receptor recycling and degradation. Research into SNX25 function contributes to a broader understanding of vesicular transport and may have relevance to disease contexts involving aberrant receptor signaling.

SNX25 Mouse mAb (Ab351394) provides researchers with a highly specific monoclonal tool for detecting and studying the SNX25 protein in experimental systems. Monoclonal antibodies offer the advantage of batch-to-batch consistency and defined epitope recognition, which are critical for reproducible results across experiments and laboratories.

Recommended Storage and Handling

Proper storage and handling of SNX25 Mouse mAb are essential to maintain antibody activity and ensure experimental reliability. The following guidelines are recommended:

• Formulation: The antibody is supplied in PBS only, without carrier proteins such as BSA or preservatives such as sodium azide. This makes the antibody directly compatible with most downstream applications without the need for dialysis or buffer exchange.
• Short-term storage: Store at 4°C for up to 2 weeks if the antibody will be used frequently within this period.
• Long-term storage: Aliquot the antibody into single-use volumes and store at −20°C or −80°C to avoid repeated freeze-thaw cycles, which can degrade antibody performance.
• Handling: Thaw aliquots gently on ice. Avoid vortexing; instead, mix by gentle pipetting. Centrifuge briefly before use if any precipitation is observed.
• Contamination prevention: Use aseptic technique and maintain a clean working environment to prevent microbial contamination of the stock.

Application Protocols and Optimization Tips

SNX25 Mouse mAb is intended for use in standard immunological and cell biological assays. While specific validated conditions should be confirmed using the current datasheet, the following general protocol guidance applies to commonly used applications:

Western Blot (WB)

1. Lyse cells or tissues in an appropriate lysis buffer containing protease inhibitors. Determine protein concentration and load 20–50 µg total protein per lane.
2. Resolve proteins by SDS-PAGE and transfer to a nitrocellulose or PVDF membrane.
3. Block membrane in 5% non-fat milk or 5% BSA in TBST for 1 hour at room temperature.
4. Incubate with SNX25 Mouse mAb at the manufacturer-recommended dilution overnight at 4°C with gentle agitation.
5. Wash three times with TBST (5 minutes each), then incubate with a compatible HRP-conjugated secondary anti-mouse antibody for 1 hour at room temperature.
6. Detect signal using ECL substrate and image using a chemiluminescence system.

Immunofluorescence (IF) / Immunocytochemistry (ICC)

1. Fix cells with 4% paraformaldehyde for 15 minutes, then permeabilize with 0.1–0.3% Triton X-100 for 10 minutes.
2. Block with 5% normal serum in PBS for 1 hour.
3. Incubate with SNX25 Mouse mAb at an appropriate dilution for 1–2 hours at room temperature or overnight at 4°C.
4. Wash with PBS and incubate with a fluorophore-conjugated secondary anti-mouse antibody.
5. Counterstain nuclei with DAPI and mount for imaging.

Optimization Note: Antibody dilutions, incubation times, and blocking conditions may require optimization depending on the specific cell line, tissue type, or detection platform used. Always include appropriate positive and negative controls.

Quality Control and Troubleshooting

Ensuring the quality of your experimental results when using SNX25 Mouse mAb requires attention to common sources of variability and failure. The following troubleshooting guide addresses frequently encountered issues:

Problem	Possible Cause	Recommended Action
No signal detected	Antibody concentration too low; antigen lost during processing	Increase antibody concentration; verify protein extraction efficiency; confirm target expression in the cell/tissue model used
High background	Insufficient blocking; secondary antibody cross-reactivity	Extend blocking time; optimize secondary antibody dilution; increase washing stringency
Multiple non-specific bands (WB)	Over-loading; cross-reactivity	Reduce sample loading; verify antibody lot performance with a positive control lysate
Weak or variable signal	Freeze-thaw degradation; suboptimal incubation conditions	Use freshly thawed aliquots; optimize incubation temperature and duration

Always verify that SNX25 is expressed in the experimental model system before concluding that the antibody has failed. Use positive control cell lines or tissues known to express SNX25.

Regulatory, Safety and Ordering Information

SNX25 Mouse mAb (Ab351394) is supplied by Aladdin Scientific as a research-use-only (RUO) reagent. It is not intended for diagnostic, therapeutic, or clinical use in humans or animals.

Safety: This product is formulated in PBS and does not contain hazardous preservatives such as sodium azide at concentrations requiring special disposal procedures. Nonetheless, standard laboratory safety practices apply: wear appropriate personal protective equipment (PPE), including gloves and eye protection, when handling. Dispose of biological materials in accordance with local institutional and regulatory guidelines.

Ordering: SNX25 Mouse mAb is available under SKU Ab351394. For bulk quantities, custom formulations, or special requirements, contact Aladdin Scientific customer service. Reference catalog number absea #KC-31961 when inquiring about cross-reference sourcing.

Returns and Quality Guarantee: If you experience quality issues with this product, please contact Aladdin Scientific within the applicable return window as outlined in our Returns and Refunds policy. Provide the lot number, storage conditions, and a description of the issue for fastest resolution.

Frequently asked questions

What is SNX25 and why is it scientifically important?

SNX25 (Sorting Nexin 25) is a member of the sorting nexin protein family, characterized by a PX domain that facilitates binding to phosphoinositides on endosomal membranes. It plays roles in intracellular membrane trafficking and endosomal sorting, making it relevant to studies of receptor regulation, vesicular transport, and potentially disease-associated signaling pathways.

Why is SNX25 Mouse mAb supplied in PBS only, and does this affect downstream applications?

The antibody is formulated in PBS without carrier proteins (e.g., BSA) or preservatives (e.g., sodium azide) to maximize compatibility with downstream applications, including enzyme-based assays where BSA could compete and azide-based preservatives could inhibit HRP activity. This formulation is directly ready for use and does not require additional buffer exchange steps.

How should I determine the optimal working dilution for SNX25 Mouse mAb in my specific assay?

Optimal dilution must be empirically determined for each application, cell/tissue type, and detection system. Start with the dilution range recommended on the product datasheet and perform a titration experiment. Include positive controls (cell lines or tissues known to express SNX25) and negative controls to define specific signal versus background.

Is SNX25 Mouse mAb validated for immunoprecipitation (IP)?

Validated applications for this antibody should be confirmed by referring to the current product datasheet for Ab351394. Not all monoclonal antibodies are suitable for immunoprecipitation, as IP requires recognition of native epitopes under non-denaturing conditions. If IP is your intended use, contact Aladdin Scientific technical support for guidance.

What secondary antibody should I use with SNX25 Mouse mAb?

Since SNX25 Mouse mAb is raised in mouse, you should use an anti-mouse secondary antibody appropriate for your detection method (e.g., HRP-conjugated anti-mouse IgG for WB, or fluorophore-conjugated anti-mouse IgG for IF/ICC). Ensure the secondary antibody is validated for use against mouse IgG and is matched to your detection platform.

References

1. Cullen PJ, Korswagen HC. Sorting nexins provide diversity for retromer-dependent trafficking events. Nat Cell Biol. 2012;14(1):29-37. doi:10.1038/ncb2374
2. Teasdale RD, Collins BM. Insights into the PX (phox-homology) domain and SNX (sorting nexin) protein families: structures, functions and roles in disease. Biochem J. 2012;441(1):39-59. doi:10.1042/BJ20111226
3. Worby CA, Dixon JE. Sorting out the cellular functions of sorting nexins. Nat Rev Mol Cell Biol. 2002;3(12):919-931. doi:10.1038/nrm974

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