Product Overview and Target Background

ARHGAP25 (Rho GTPase Activating Protein 25) is a member of the RhoGAP family of proteins, which function as negative regulators of Rho family GTPases by stimulating their intrinsic GTPase activity. Rho GTPases play central roles in actin cytoskeleton organization, cell migration, and immune cell function. ARHGAP25 has been reported to be particularly relevant in hematopoietic cells, where it regulates phagocytosis and cell motility.

The ARHGAP25 Mouse Monoclonal Antibody (SKU: Ab351156) is produced in mouse and is formulated in phosphate-buffered saline (PBS) without additional carriers or preservatives. Researchers should verify specific application compatibility — such as Western blotting (WB), immunohistochemistry (IHC), immunofluorescence (IF), or flow cytometry (FC) — against validated data for their experimental system, as the source material does not specify validated applications explicitly.

Storage, Handling, and Reconstitution

Proper storage and handling are critical to maintaining antibody activity and specificity. Follow the guidelines below for ARHGAP25 Mouse mAb (Ab351156):

• Short-term storage (up to 1 month): Store at 4 °C. Avoid repeated freeze-thaw cycles.
• Long-term storage: Store at −20 °C or −80 °C in small aliquots to minimize freeze-thaw degradation.
• Formulation: The antibody is supplied in PBS only. If your application requires a stabilizer such as glycerol or BSA, consider adding them according to standard laboratory practice; consult Aladdin Scientific technical support for guidance.
• Avoid: Exposure to repeated freeze-thaw cycles, prolonged exposure to light, and contamination. Use sterile technique when aliquoting.

Note: Specific storage temperature and shelf-life data are not detailed in the provided source material. Users should refer to the product datasheet or contact Aladdin Scientific for confirmed storage conditions.

Recommended Protocol and Dilution Guidelines

While application-specific validated dilutions for Ab351156 are not explicitly stated in the provided source material, the following general guidelines apply to mouse monoclonal antibodies and should be treated as starting points, to be optimized empirically in your laboratory:

• Western Blotting (WB): A typical starting dilution range is 1:500–1:2000. Use standard SDS-PAGE and transfer protocols. Block with 5% non-fat milk or BSA in TBST.
• Immunohistochemistry (IHC): Starting dilution of 1:50–1:200 is common. Antigen retrieval (heat-mediated, citrate buffer pH 6.0) may be required.
• Immunofluorescence (IF): Suggested starting dilution of 1:100–1:500. Use an appropriate secondary antibody conjugated to a fluorophore.
• Flow Cytometry (FC): Titrate from 1–5 µg per 10⁶ cells; optimize based on signal-to-noise ratio.

Always include appropriate positive and negative controls. Refer to the official Aladdin Scientific product datasheet for application-validated dilutions and conditions specific to Ab351156.

Troubleshooting Common Issues

If unexpected results are observed when using ARHGAP25 Mouse mAb (Ab351156), consider the following troubleshooting steps:

• No signal / Weak signal: Verify target expression in your cell or tissue type. Check antibody concentration and optimize dilution. Ensure proper antigen retrieval for IHC applications. Confirm secondary antibody compatibility.
• High background: Increase blocking time or concentration. Optimize antibody dilution. Reduce primary antibody incubation time or temperature. Ensure washing steps are thorough.
• Non-specific bands (WB): Optimize antibody dilution. Use freshly prepared samples and avoid protein degradation. Verify gel percentage is appropriate for the molecular weight of ARHGAP25.
• Inconsistent results between experiments: Aliquot the antibody to prevent freeze-thaw cycles. Standardize sample preparation and lysis buffers across experiments.

Contact Aladdin Scientific technical support at any stage for application-specific guidance.