SOX8 Mouse mAb: Applications, Usage & Best Practices

SOX8 Mouse mAb is a highly specific monoclonal antibody raised in mouse against the SOX8 transcription factor, designed to enable reliable detection and characterization of SOX8 protein in a range of immunological and biochemical applications.

Product SKU

Ab349648

Antibody Type

Mouse Monoclonal Antibody (mAb)

Target Protein

SOX8 (SRY-box transcription factor 8)

Host Species

Mouse

Recommended Applications

WB, IHC, IF (verify specific validated applications on product datasheet)

Storage Recommendation

Store at -20°C; avoid repeated freeze-thaw cycles (confirm on datasheet)

Background: SOX8 and Its Biological Role

SOX8 (SRY-box transcription factor 8) is a member of the SOX family of transcription factors, which are characterized by a highly conserved high-mobility group (HMG) box DNA-binding domain. SOX family members play critical roles in regulating development, cell fate determination, and differentiation across a wide range of tissues.

SOX8 is expressed in multiple tissues including the central nervous system, gonads, and various other organs during development. It has been implicated in neural development, Sertoli cell function, and has emerging roles in cancer biology, where altered SOX8 expression has been associated with tumor progression and metastasis in several cancer types.

Accurate detection of SOX8 protein expression is essential for studies investigating developmental biology, stem cell research, and oncology. The SOX8 Mouse mAb (Ab349648) provides a reliable tool for these investigations.

Recommended Protocol and Application Guidelines

Western Blot (WB): Prepare cell or tissue lysates using standard RIPA or equivalent lysis buffer supplemented with protease inhibitors. Resolve proteins by SDS-PAGE, transfer to PVDF or nitrocellulose membrane, and block with 5% non-fat milk or BSA in TBST for 1 hour at room temperature. Incubate with SOX8 Mouse mAb at the manufacturer-recommended dilution (refer to product datasheet for validated dilution range) overnight at 4°C. Wash three times with TBST, then incubate with an appropriate HRP-conjugated anti-mouse secondary antibody. Detect signal by chemiluminescence.

Immunohistochemistry (IHC): Use formalin-fixed, paraffin-embedded (FFPE) tissue sections. Perform antigen retrieval using citrate buffer (pH 6.0) or EDTA buffer (pH 9.0) as appropriate. Block endogenous peroxidase and non-specific binding before incubating with the primary antibody at the recommended dilution. Apply appropriate secondary detection system and counterstain with hematoxylin.

Immunofluorescence (IF): Fix cells with 4% paraformaldehyde and permeabilize with 0.1–0.5% Triton X-100. Block with 5% normal serum, then incubate with SOX8 Mouse mAb at the recommended dilution. Detect with a fluorophore-conjugated anti-mouse secondary antibody and counterstain nuclei with DAPI if desired.

Note: Always consult the official product datasheet for Ab349648 for validated dilutions, positive controls, and species reactivity prior to use.

Storage, Handling, and Quality Considerations

Storage: SOX8 Mouse mAb should be stored according to the conditions specified on the product datasheet, typically at -20°C for long-term storage. Aliquoting the antibody upon first use is strongly recommended to minimize the number of freeze-thaw cycles, which can compromise antibody integrity and performance.

Handling: Keep the antibody on ice during experimental setup. Avoid prolonged exposure to light if the antibody is conjugated to a fluorophore. Centrifuge briefly before opening to collect any material from the tube cap or walls.

Quality Control: Each lot of SOX8 Mouse mAb undergoes quality control testing by the manufacturer. Lot-specific data, including validated applications and tested dilutions, are provided on the datasheet accompanying the product. Researchers should validate antibody performance in their specific experimental system, as results can vary depending on sample preparation, tissue type, and detection method.

Expiry: Use the antibody within the expiration date stated on the vial label. Degraded antibody may produce high background, loss of specific signal, or inconsistent results.

Troubleshooting Common Issues

No signal or weak signal: Verify that the antibody concentration and dilution are appropriate for your application. Ensure antigen retrieval (for IHC) was performed correctly. Check that the secondary antibody is compatible with the mouse host species of the primary antibody. Confirm that the target protein is expressed in the chosen sample.

High background: Increase blocking time or concentration. Optimize antibody dilution by titration. Increase the number and duration of wash steps. Ensure that the secondary antibody does not cross-react with the sample species.

Non-specific bands (WB): Increase antibody dilution and optimize blocking conditions. Use fresh lysis buffer with appropriate protease inhibitors to prevent protein degradation. Run a negative control (cell line or tissue known to lack SOX8 expression) to confirm band specificity.

Inconsistent results between experiments: Standardize sample preparation protocols. Aliquot the antibody to avoid repeated freeze-thaw cycles. Verify consistent antigen retrieval conditions (time, temperature, buffer pH) between experiments.

Frequently asked questions

What species does SOX8 Mouse mAb (Ab349648) react with?

Species reactivity depends on the specific lot and is validated by the manufacturer. Please refer to the official Ab349648 product datasheet for confirmed reactive species, as this information is lot-specific and subject to change.

Which applications is SOX8 Mouse mAb validated for?

SOX8 Mouse mAb is intended for use in immunological and biochemical detection applications such as Western Blot (WB), Immunohistochemistry (IHC), and Immunofluorescence (IF). The specific validated applications and recommended working dilutions for Ab349648 are detailed in the product datasheet. Always confirm validated applications before experimental use.

How should I store and handle SOX8 Mouse mAb to maintain its activity?

Store SOX8 Mouse mAb at -20°C for long-term stability, as typically recommended for monoclonal antibodies. Upon first use, aliquot the antibody into single-use volumes to avoid repeated freeze-thaw cycles, which can degrade antibody performance. Always keep the antibody on ice during experiments and refer to the product datasheet for storage conditions specific to Ab349648.

What positive control should I use when working with SOX8 Mouse mAb?

A suitable positive control is a cell line or tissue known to express SOX8 endogenously, such as certain neural or testicular tissue samples. Refer to the Ab349648 product datasheet for manufacturer-recommended positive controls, as validated controls are lot-specific.

References

1. Wegner M. From head to toes: the multiple facets of Sox proteins. Nucleic Acids Research. 1999;27(6):1409-1420. doi:10.1093/nar/27.6.1409
2. Sock E, Wegner M. Transcriptional control of myelination and remyelination. Glia. 2019;67(11):2091-2103.
3. Aladdin Scientific. Ab349648 SOX8 Mouse mAb Product Datasheet. Aladdin Scientific; [current lot].

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