Sc 393998

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Sc-393998 is a laboratory instrument designed for protein purification. It utilizes chromatographic techniques to isolate and concentrate target proteins from complex biological samples. The core function of this equipment is to enable the efficient separation and purification of proteins for various research and analytical applications.

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Lab products found in correlation

5 protocols using sc 393998

Molecular Mechanisms of Nur77 Regulation

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NK (purity, >98%) was acquired from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China) (495–31–8) (250 mg administered). Lipopolysaccharide (LPS, 297–473–0) was acquired from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against α-SMA (bs-10196R) and collagen type I (collagen-I) (bs-0578R) were obtained from Bioss (Woburn, MA, USA). Antibodies against PPARα (sc-398394), SREBP-1 (sc-365513), caspase-1 (sc-56036), IL-23 (sc-271279), ASC (sc-514414), Lipin-1 (sc-376874), Nur77 (sc-365113), NLRP3 (ab4207), P2X7r (sc-514962), and IL-1R1 (sc-393998) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against NLRP3 (ab4207), Lipin-1 (ab181389), P2X7r (ab307718), MPO (ab208670), and GAPDH (ab8245) were obtained from Abcam (Cambridge, MA, USA). Csn-B (a Nur77 agonist, 321661–62–5) was purchased from Beyotime Biotechnology (Shanghai, CN, USA). rNur77 (ab152448) was purchased from Abcam (Cambridge, MA, USA).

Song J., Qin B.F., Zhang J.J., Feng Q.Y., Liu G.C., Zhao G.Y, & Sun H.M. (2024). Regulation of the Nur77-P2X7r Signaling Pathway by Nodakenin: A Potential Protective Function against Alcoholic Liver Disease. Molecules, 29(5), 1078.

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Immunocytochemical Analysis of synMSCs

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Immunocytochemical analyses were performed as described previously [17 (link)]. Briefly, synMSCs were seeded at 5000–10000 cells on 8-well chamber slides (Corning#354118, New York, NY, USA) and cultured for 5 days. Cells were fixed in 4% paraformaldehyde (Wako#162-16065, Tokyo, Japan) and permeabilized in methanol (Wako#137-01823, Tokyo, Japan). Cells were incubated with CD121a antibodies (Santa Cruz#sc-393998, 1:100) overnight at 4 °C and anti-mouse IgG antibodies (Thermo Fisher#A11001, 1:250) at 20–25 °C for 1 h. To determine the localization of the cell membrane, antibodies against Na⁺/K⁺ ATPases (Abcam#ab70620, 1:500) were co-stained with the CD121a antibody. Images were obtained using a BX63 automated fluorescence microscope (Olympus, Tokyo, Japan). Analysis was performed using ImageJ software (NIH, USA).

Tang G., Asou Y., Matsumura E., Nakagawa Y., Miyatake K., Katagiri H., Nakamura T., Koga H., Komori K., Sekiya I., Ezura Y, & Tsuji K. (2022). Short cytoplasmic isoform of IL1R1/CD121a mediates IL1β induced proliferation of synovium-derived mesenchymal stem/stromal cells through ERK1/2 pathway. Heliyon, 8(5), e09476.

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Quantification of Immune Receptors

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Cell protein homogenates (25 μg) were separated by electrophoresis in 12% polyacrylamide gels, transferred to nitrocellulose membranes, and blocked overnight with 5% nonfat dry milk. The membranes were washed and incubated in the presence of the following monoclonal antibodies: anti-IL-1R1, anti-IL-1R2, anti-TNFR1, anti-TNFR2, anti-VDR (sc-393998, sc-376247, sc-8436, sc-393614, and sc-13133, respectively; Santa Cruz Biotechnology, CA, USA), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH, MAB374, Millipore, Milford, MA, USA) overnight at 4°C. Membranes were incubated in the presence of secondary antibody conjugated with horseradish peroxidase (sc-2031, Santa Cruz Biotechnology) for 2 hours at room temperature. The immunoblots were visualized by chemiluminescence using ECL Plus (Amersham Pharmacia, UK).

Martínez-Reza I., Díaz L., Barrera D., Segovia-Mendoza M., Pedraza-Sánchez S., Soca-Chafre G., Larrea F, & García-Becerra R. (2019). Calcitriol Inhibits the Proliferation of Triple-Negative Breast Cancer Cells through a Mechanism Involving the Proinflammatory Cytokines IL-1β and TNF-α. Journal of Immunology Research, 2019, 6384278.

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Immunohistochemical Analysis of Inflammatory Receptors in Bone Marrow

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After the mice (n = 3 each group) were euthanized after 24 h reperfusion, the femur, tibia, and fibula were harvested for histological studies. Immunohistochemical staining was used to detect the expression of IL-1RI, IL-1RII, and IL-1Ra in the BM. Paraffin sections were deparaffinized and rehydrated, then deposited and stored at 4 °C overnight before being incubated with primary antibodies IL-1RI (1:50, sc-393998, Santa Cruz, USA), IL-1RII (1:50, sc-376247, Inc), IL-1RII (1:50, sc-376247, Santa Cruz, USA), and IL-1Ra (1:50, sc-374084, Santa Cruz, USA). The next day, the corresponding secondary antibody was applied at 20 to 25 °C for about 50 min, followed by the prepared 3,3'-diaminobenzidine (DAB) substrate and hematoxylin. The results were evaluated by two qualified pathologists. Motic Images Plus version 2.0 system (Motic China Group Co, Xiamen, China) was used for image acquisition and the ImageJ analysis system was quantified.

Tan Y., Bao X., Li Y., Song G., Lu H., Sun X., Gu R., Kang L, & Xu B. (2023). Colchicine Attenuates Microvascular Obstruction after Myocardial Ischemia-Reperfusion Injury by Inhibiting the Proliferation of Neutrophil in Bone Marrow. Cardiovascular drugs and therapy.

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Protein Expression Analysis in Cumulus Cells

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Collected cumulus cells and oocytes were lysed with 2× loading buffer as western blot samples. A 12–15% SDS gel was used to separate proteins and then transfer the protein to a nitrocellulose membrane. After being sealed in non-fat milk for 1 h at 37 °C, the membrane containing protein was incubated with antibodies of IL1A (1:300; sc-12741, Santa Cruz, Dallas, TX, USA), IL1B (1:500; ab234437, Abcam, Cambridge, UK), IL1R1 (1:300; sc-393998, Santa Cruz), REIA (1:500; WL01980, Wanleibio, Shenyang, China), NFKB1 (1:500; WL01866, Wanleibio), and tubulin (1:1000; ab6046, Abcam). After washing in TBST, the membrane was incubated at room temperature with a homologous secondary antibody for 2 h. ECL Plus (GE, Piscataway, NJ, USA) and the Tanon 3900 chemiluminescence imaging system (Tanon, Beijing, China) were used to color and observe protein bands, respectively.

Wen X., Yang Q., Sun D., Jiang Z.Y., Wang T., Liu H.R., Han Z., Wang L, & Liang C.G. (2023). Cumulus Cells Accelerate Postovulatory Oocyte Aging through IL1–IL1R1 Interaction in Mice. International Journal of Molecular Sciences, 24(4), 3530.

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