Sc 8992

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Sc-8992 is a laboratory equipment product manufactured by Santa Cruz Biotechnology. It is designed for general scientific applications within a laboratory setting. The core function of Sc-8992 is to provide a precise and reliable platform for conducting various experiments and analyses.

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Lab products found in correlation

4 protocols using sc 8992

Immunohistochemical Analysis of DRG Neurons

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Mice were anesthetized with 80 mg/kg of ketamine and 10 mg/kg of xylazine before transcardial perfusion with 30 ml 0.1 M PBS (pH 7.4) followed by 100 ml of 4% PFA in PBS. DRGs were embedded in optimal cutting temperature compound (OCT; Tissue-Tek, VWR International) and frozen at -80°C; 10- to 20-μm sections were cut using a cryostat and thaw-mounted on SuperFrost Plus slides (Fisher Scientific), then stored at -20°C until use. After drying at RT, slides were rinsed with 0.1 M PBS and overlaid with blocking solution for 1 h. Sections were subsequently washed and incubated overnight with the primary antibody and appropriate secondary antibodies. We used the following primary and secondary antibodies: GR: 1:200, #PA1-511A, Thermo Fisher, RRID: AB_2236340 or sc-8992, Santa Cruz Biotechnology, RRID: AB_2155784; SCG10, 1:500, Novus Biotechnologies anti-stathmin-2, NBP1-49467, RRID: AB_10011568; ATF3, 1:500, sc-188, Santa Cruz, RRID: AB_2258513; NFH, 1:1000; Aves Labs, RRID: AB_2313552; 555 or 488-conjugated goat anti-rabbit IgG antibody, 1:500 #A11035, Invitrogen. Hoechst or DAPI was applied in the final rinse to visualize nuclei.

Lerch J.K., Alexander J.K., Madalena K.M., Motti D., Quach T., Dhamija A., Zha A., Gensel J.C., Webster Marketon J., Lemmon V.P., Bixby J.L, & Popovich P.G. (2017). Stress Increases Peripheral Axon Growth and Regeneration through Glucocorticoid Receptor-Dependent Transcriptional Programs. eNeuro, 4(4), ENEURO.0246-17.2017.

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Western Blot Analysis of Protein Markers

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Cells were lysed in RIPA buffer and protein was loaded on a SDS–PAGE gel and subsequently transferred to a nitrocellulose membrane. Membranes were incubated with the primary antibody overnight at 4 °C and with the secondary antibody for 1 h at room temperature. The following antibodies were used: anti-GR (1:500, sc-8992, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-GAPDH (1:10 000, Chemicon, Temecula, CA, USA), anti-β-actin (1:5000, Sigma), anti-Bcl-xL (1:1000, sc-23958, Santa Cruz Biotechnology), and anti-Bcl-2 (1:1000, sc-7382, Santa Cruz Biotechnology).

Kroon J., Puhr M., Buijs J.T., van der Horst G., Hemmer D.M., Marijt K.A., Hwang M.S., Masood M., Grimm S., Storm G., Metselaar J.M., Meijer O.C., Culig Z, & van der Pluijm G. (2016). Glucocorticoid receptor antagonism reverts docetaxel resistance in human prostate cancer. Endocrine-Related Cancer, 23(1), 35-45.

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ChIP Analysis of GR Regulation in BMDMs

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ChIP analysis of wild-type and GR^dim BMDMs was performed 4 h after stimulation with LPS (100 ng ml⁻¹) and Dex (10⁻⁶ M). ChIP assays were performed as described elsewhere70 (link). The anti-GR antibody (sc-8992) was obtained from Santa Cruz Biotechnology. The following primers were used: SphK1 forward 5′- GCTGCTGATGTGAAGGATAC -3′ and SphK1 reverse 5′- GCTGCTGATGTGAAGGATAC -3′. Foxl2 was selected as a negative control and the following primers were used: forward 5′- GCTGGCAGAATAGCATCCG -3′ and reverse 5′- TGATGAAGCACTCGTTGAGGC -5′. The fold enrichment over immunoglobulin G was calculated. Putative GC-response element-binding sites were identified by Genomatix software.

Vettorazzi S., Bode C., Dejager L., Frappart L., Shelest E., Klaßen C., Tasdogan A., Reichardt H.M., Libert C., Schneider M., Weih F., Henriette Uhlenhaut N., David J.P., Gräler M., Kleiman A, & Tuckermann J.P. (2015). Glucocorticoids limit acute lung inflammation in concert with inflammatory stimuli by induction of SphK1. Nature Communications, 6, 7796.

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Protein Expression Analysis by Western Blot

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Proteins concentration was measured using Lowry method (Lowry et al. 1951 ) and 60 µg of proteins were subjected to electrophoresis on 7.5% SDS-PAGE. After transfer to PVDF membrane (Immobilon-FL membrane, Millipore), using a Transblot system (Bio-Rad Laboratories), the unbound sites were blocked (1 h with 2% non-fat dry milk) and the membranes were probed with specific primary rabbit polyclonal antibodies for anti-phospho-p38 Tyr 182 (1:750) (sc-7975-R, Santa Cruz Biotechnology), anti-p38 MAPK (1:500) (sc-535, Santa Cruz Biotechnology), anti-GR (1:500) (sc-8992, Santa Cruz Biotechnology), anti-11BHSD1 (1:1000) (ab393364, Abcam), anti-H6PDH (1:1000) (sc-67394, Santa Cruz Biotechnology), anti-SREBP1c (1:500) (sc-366, Santa Cruz Biotechnology) and mouse monoclonal anti-PPARG (1:1000) (sc-7273, Santa Cruz Biotechnology). Monoclonal mouse anti-B-actin antibody (1:10,000) (AC-15, Sigma-Aldrich) was used as a loading control. Blots were developed with secondary ECL anti-rabbit IgG HRP-linked whole antibody (1:10,000) (Amersham Pharmacia Biotech) or with anti-mouse IgG HRP-linked whole antibody (1:20,000) (ab97046, Abcam). Signal was developed using enhanced chemiluminescence Downloaded from Bioscientifica.com at 09/07/2024 04:13:17AM via free access (ECL) and densitometry of protein bands on X-ray films (Kodak, Rochester, USA) was performed by Image J software (NIH).

Gligorovska L., Bursać B., Kovačević S., Veličković N., Matić G, & Djordjevic A. (2019). Mif deficiency promotes adiposity in fructose-fed mice. The Journal of endocrinology, 240(2).

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