Anti 11β hsd2 antibody

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The Anti-11β-HSD2 antibody is a laboratory-grade reagent designed for use in various research applications. It specifically targets the 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) enzyme, which plays a role in the metabolism of glucocorticoids. This antibody can be utilized in techniques such as Western blotting, immunohistochemistry, and immunoprecipitation to detect and study the 11β-HSD2 protein. Its core function is to provide researchers with a tool for analyzing the expression and localization of 11β-HSD2 in biological samples.

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Lab products found in correlation

Antibody diluent, by Agilent Technologies (1 mentions) Protein block serum free, by Agilent Technologies (1 mentions) Vectashield hardset mounting medium, by Vector Laboratories (1 mentions) Chamber slide, by BD (1 mentions) Formaldehyde neutral buffer solution, by Merck Group (1 mentions) Envision kit hrp dab, by Agilent Technologies (1 mentions) Bz 9000, by Keyence (1 mentions) Chemiluminescent detection system, by Thermo Fisher Scientific (1 mentions) Nitrocellulose membrane, by Merck Group (1 mentions) Ripa lysis buffer, by Merck Group (1 mentions) Anti β actin antibody, by Merck Group (1 mentions) Protease inhibitor, by Roche (1 mentions)

Close spelling variants of this product

11β-HSD2

3 protocols using anti 11β hsd2 antibody

Immunofluorescence Assay for 11β-HSD2 in Prostate Cancer Cells

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LAPC4 cells treated with Enz or vehicle for 23 days were seeded into chamber slides (BD Biosciences) coated with poly-DL-ornithine at 60% confluence. After overnight culture, cells were washed with PBS and fixed with ice cold methanol for 15 min and the methanol was washed with PBS. Before applying primary antibodies, nonspecific binding sites were blocked with blocking buffer (Protein Block Serum Free, Dako). Anti-11β-HSD2 antibody (Santa Cruz), diluted at 1:300 with Antibody Diluent (Dako), was applied for incubation overnight at 4°C. After being rinsed with PBS, the slides were probed with Alexa Fluor 594 conjugated secondary antibody (goat anti-rabbit, Thermo Scientific) for 45 min at room temperature. VECTASHIELD HardSet Mounting Medium (Vector Laboratories) was used to mount the slides and counterstain the nucleus with DAPI.

Li J., Alyamani M., Zhang A., Chang K.H., Berk M., Li Z., Zhu Z., Petro M., Magi-Galluzzi C., Taplin M.E., Garcia J.A., Courtney K., Klein E.A, & Sharifi N. (2017). Aberrant corticosteroid metabolism in tumor cells enables GR takeover in enzalutamide resistant prostate cancer. eLife, 6, e20183.

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Histological Examination of Maxillary Bones

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Another set of experiments was performed for histological examinations. After euthanasia, the rats were perfused with 10 % formaldehyde neutral buffer solution (Sigma-Aldrich, St. Louis, MO, USA). Then the maxillary bones were excised, and fixed in 10 % formaldehyde neutral buffer solution at 4 °C for 3 days. The bones were decalcified in a rapid decalcification solution, K-CX (Falma, Tokyo, Japan), at 4 °C for 24 h, followed by conventional dehydration and paraffin embedding. After cutting into 5 μm-thick sections, the specimens were deparaffinized and then stained with hematoxylin-eosin (HE) or immunostained with either anti-11β-HSD1 antibody (Bioss, Woburn, MA, USA) or anti-11β-HSD2 antibody (Santa Cruz Biotechnology, Dallas, TX, USA) using Envision + kit/HRP (DAB) (Dako, Glostrup, Denmark). Images were obtained using an all-in-one microscope, BZ-9000 (Keyence, Osaka, Japan).

Nakata T., Umeda M., Masuzaki H, & Sawai H. (2016). The expression of 11β-hydroxysteroid dehydrogenase type 1 is increased in experimental periodontitis in rats. BMC Oral Health, 16, 108.

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Steroid-Induced Gene and Protein Expression

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Forward: 5’-CGAGGGCACCTGGAAAAC-3’;
Reverse: 5’-CACATTCCCCCGGATATGA-3’.
For steroid-treated cells, each mRNA transcript was quantitated by normalizing the sample values to RPLP0 and to vehicle-treated cells. All gene expression studies were repeated in at least three independent experiments.
For protein analysis, immunoblots were performed as described previously (6 ). Briefly, total cellular protein was extracted with ice-cold RIPA lysis buffer (Sigma-Aldrich) containing protease inhibitors (Roche). Protein concentration was determined using a BCA protein assay (Pierce Protein Research Products, Thermo Scientific). Protein, 30–50 μg, was separated by 8% SDS-PAGE and then transferred to a nitrocellulose membrane (Millipore). After incubating with the anti-11β-HSD2 antibody (Santa Cruz; 1:3000), anti-H6PD antibody (Origene; 1:6000) overnight at 4°C, the appropriate secondary antibody was incubated for 1 hour at room temperature. A chemiluminescent detection system (Thermo Scientific) was used to detect the bands with peroxidase activity. An anti-β-actin antibody (Sigma-Aldrich; 1:5000) was used as a control for sample loading.

Jianneng L.i., Berk M., Alyamani M., Sabharwal N., Goins C., Alvarado J., Baratchian M., Zhu Z., Stauffer S., Klein E.A, & Sharifi N. (2021). Hexose-6-phosphate dehydrogenase blockade reverses prostate cancer drug resistance in xenograft models by glucocorticoid inactivation. Science translational medicine, 13(595), eabe8226.

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