Description

Nori Human SOX9 ELISA Kit Summary

Alternative names for SOX9: Transcription factor SOX-9, SOX-9

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Transcription factor SOX-9 is a protein that is encoded by the SOX9 gene. SOX-9 recognizes the sequence CCTTGAG along with other members of the HMG-box class DNA-binding proteins. It is expressed by proliferating but not hypertrophic chondrocytes that is essential for differentiation of precursor cells into chondrocytes and, with steroidogenic factor 1. SOX9 regulates transcription of the anti-Müllerian hormone (AMH) gene. SOX-9 plays a pivotal role in male sexual development; by working with Sf1, SOX-9 can produce AMH in Sertoli cells to inhibit the creation of a female reproductive system.^[1] Activation of FGF9 by SOX-9 starts vital processes in male development, such as the creation of testis cords and the multiplication of Sertoli cells.^[2] The association of SOX-9 and Dax1 actually creates Sertoli cells, another vital process in male development.^[3] In the brain development, its murine ortholog Sox-9 induces the expression of Wwp1, Wwp2, and miR-140 to regulate cortical plate entry of newly born nerve cells, and regulate axon branching and axon formation in cortical neurons.^[4] SOX-9 is a target of the Notch signaling pathway, as well as the Hedgehog pathway,^[5] and plays a role in the regulation of neural stem cell fate. In vivo and in vitro studies show that SOX-9 negatively regulates neurogenesis and positively regulates gliogenesis and stem cell survival.^[6]

The Sox9 protein has been implicated in both initiation and progression of multiple solid tumors. Its role as a master regulator of morphogenesis during human development makes it an ideal candidate for perturbation in malignant tissues. Specifically, Sox9 appears to induce invasiveness and therapy-resistance in prostate,^[7] colorectal,^[8] breast^[9] and other cancers, and therefore promotes lethal metastasis. Many of these oncogenic effects of Sox9 appear dose dependent.

References

1. De Santa Barbara P, et al. (1998). Molecular and Cellular Biology. 18 (11): 6653–65.
2. Kim Y, et al. (2006). PLOS Biology. 4 (6): e187. doi:10.1371/journal.pbio.0040187.
3. Bouma GJ, et al. (2005). Development. 132 (13): 3045–54. doi:10.1242/dev.01890.
4. Ambrozkiewicz MC, et al. (2018). Neuron. 100 (5): 1097–1115.e15.
5. Place E, et al. (2022) Front. Neurosci. 16:855288. doi: 10.3389/fnins.2022.855288
6. Vogel, Julia K., et al. (2014). Genes & Diseases. 1 (2): 149–161.
7. Nouri M, et al. (2020). Clinical Cancer Research. 26 (7): 1678–1689.
8. Prévostel C, Blache P (2017). European Journal of Cancer. 86: 150–157.
9. Grimm D, et al. (2019). Seminars in Cancer Biology. 67 (Pt 1): 122–153.

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