S 2012 50

Manufactured by Vector Laboratories

Sourced in United States

The S-2012-50 is a laboratory equipment product from Vector Laboratories. It serves as a core function for various laboratory applications.

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Lab products found in correlation

Ab108930, by Abcam (1 mentions) Hoechst, by Merck Group (1 mentions) Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions) Af1865, by R&D Systems (1 mentions) Af457, by R&D Systems (1 mentions) Af2535, by R&D Systems (1 mentions) Kma 0100 00a, by CellPath (1 mentions) Normal horse serum, by Vector Laboratories (1 mentions) Dv2004g1, by Biocare Medical (1 mentions) Pk 7200, by Vector Laboratories (1 mentions) Sc 7294, by Santa Cruz Biotechnology (1 mentions) D21490, by Thermo Fisher Scientific (1 mentions) Diamidino 2 pehnylindole dapi, by Thermo Fisher Scientific (1 mentions) Lsm 700 confocal laser scanning microscope, by Zeiss (1 mentions) Sk 4100, by Vector Laboratories (1 mentions) Aperio cs2 slide scanner, by Leica (1 mentions)

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S-2012 (13 citations)

4 protocols using s 2012 50

Multiplexed Immunofluorescence Staining of Frozen Tissue

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5 μm sections of frozen tissue embedded in OCT (KMA-0100-00A, CellPath) were fixed in ice-cold acetone, and non-specific binding blocked with 2.5% normal horse serum (S-2012-50, VectorLabs). Sections were incubated in primary antibody overnight at 4°C, washed with PBS, incubated in secondary antibody, washed again, counterstained with Hoechst (14533, Sigma) and then mounted in ProLong Gold Antifade mountant (P36930, ThermoFisher Scientific). Primary antibodies were used as follows: anti-TNC (ab108930, abcam) 1 μg/mL, anti-CD3 (GA50361-2, Agilent) 2 μg/mL, anti-GZMB (AF1865, R&D Systems) 0.4 μg/mL, anti-CD8 (42-0081-82, ThermoFisher) 1 mg/mL, anti-Lumican (AF2745, R&D Systems) 0.4 μg/mL, anti-LYVE-1 (14-0443-82, ThermoFisher), anti-CCL21 (AF457, R&D Systems) 5 μg/mL, anti-ZEB1 (HPAO27524, Atlas Antibodies) 0.2 μg/mL, anti-CD206 (AF2535, R&D Systems) 0.3 mg/mL, and anti-MHC class II (NBP1-43312, Novus Biologicals) 20 μg/mL. Sections were imaged using a Zeiss Axioscan.Z1 Slide Scanner and analysed using Zen software Blue edition. Immune cells were counted per high power field of view, and an average of 10 fields of view was calculated per section.

Pires A., Greenshields-Watson A., Jones E., Smart K., Lauder S.N., Somerville M., Milutinovic S., Kendrick H., Hindley J.P., French R., Smalley M.J., Watkins W.J., Andrews R., Godkin A, & Gallimore A. (2020). Immune Remodelling of the Extracellular Matrix Drives Loss of Cancer Stem Cells and Tumor Rejection. Cancer immunology research, 8(12), 1520-1531.

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Lung Adenoma Proliferation Assay

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5 μm lung sections from urethane, iloprost, and saline groups were deparaffinized, blocked with 0.3% hydrogen peroxide, and antigen retrieval performed in boiling Diva Decloaker (Biocare Medical, DV2004G1) under pressure for 5 minutes. Sections were blocked with Background Punisher (Biocare Medical BP974M) and 2.5% Normal Horse serum (Vector Labs S-2012-50). Sections were incubated in Ki67 primary Ab (1:2000 dilution, Abcam 15580) for 1 hour at room temperature, followed by anti-rabbit universal antibody for 30 minutes and ABC reagent for 30 minutes at room temperature (Vector Laboratories PK-7200). Ki67+ nuclei were detected using Betazoid DAB chromagen kit (Vector Laboratories BDB2004). Tumor area was measured and Ki67-positive nuclei/mm² were counted in each tumor. Replicate blinded counts were conducted. The Ki67+ nuclei/mm² tumor area for each tumor was averaged by group (Dwyer-Nield et al., 2017 (link)). H&E stains were done on 5 μm lung sections from urethane, iloprost, and saline groups and were used to confirm adenoma presence and structure.

Sompel K., Dwyer-Nield L.D., Smith A.J., Elango A., Backos D.S., Zhang B., Gross J., Ternyak K., Matsuda J.L., Kopf K., Keith R.L, & Tennis M.A. (2022). Iloprost requires the Frizzled-9 receptor to prevent lung cancer. iScience, 25(6), 104442.

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Assessing NF-κB and NFATc1 Translocation

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To determine of NF-κB p65 and NFATc1 translocation, immunocytochemistry was performed. After treatment, the cells were rinsed with PBS and fixed with 4% paraformaldehyde in PBS for 10 min at room temperature. The cells were then permeabilized with 0.2% Triton X-100 in PBS for 10 min at room temperature. Nonspecific staining was blocked by 2.5% normal horse serum blocking solution (S-2012-50; Vector Laboratories, Burlingame, CA, USA) for 10 min. After blocking, the cells were incubated with primary antibodies against total NF-κB (3039; Cell Signaling) or NFATc1 (sc-7294; Santa Cruz). All antibodies were diluted 1:200 in 2.5% normal horse serum for overnight at 4 °C. Rhodamine-conjugated anti mouse (1:500; AP124R, Merk Millipore) secondary antibodies were used. The cell nuclei were stained with diamidino-2-pehnylindole (DAPI) (D21490; Thermo Fisher Scientific). The images of NF-κB and NFATc1 translocation were taken using a LSM 700 laser scanning confocal microscope (Carl zeiss, Overkochen, Germany).

Jeong S., Lee B.S., Jung S.E., Yoon Y., Song B.W., Kim I.K., Choi J.W., Kim S.W., Lee S, & Lim S. (2023). A Low Concentration of Citreoviridin Prevents Both Intracellular Calcium Deposition in Vascular Smooth Muscle Cell and Osteoclast Activation In Vitro. Molecules, 28(4), 1693.

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Cryosectioning and Histochemical Analysis of Scaffolds

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At 5 and 10 weeks, each scaffold was embedded in optimal cutting temperature liquid (OCT) and cryosectioned at 10 μm using the Kawamoto tape method [Kawamoto and Shimizu, 1986 (link)]. Slices were then fixed in 10% buffered formalin, glued to glass slides using Krazy Glue superglue, and allowed to dry completely. Calcium deposition and cell density in each of the sections was visualized through Von Kossa staining (n = 3–6). For immunohistochemical detection of osteocalcin (Santa Cruz Biotechnology, ABOC-5021) and osteopontin (Novus Biologicals, NBP1–59190), sections (n = 3–6) were incubated in proteinase K for 5 minutes followed by 3% hydrogen peroxide for 10 minutes, both at room temperature. Sections were then incubated in a blocking solution of 2.5% normal horse serum (Vector Laboratories, S-2012–50) for 30 minutes at room temperature. Osteocalcin (2 μg/ml) and osteopontin (10 μg/ml) antibodies were applied in a humidity chamber at 4°C overnight followed by a one hour room temperature incubation with R.T.U Biotinylated Secondary Antibody (Vector Laboratories, BP-1400–40). Antibody binding was visualized using a DAB chromagen reagent (Vector Laboratories, SK-4100) according to the manufacturer’s protocol. Brightfield images were acquired using a Leica Biosystems Aperio CS2 slide scanner.

Fainor M., Mahindroo S., Betz K.R., Augustin J., Smith H.E., Mauck R.L, & Gullbrand S.E. (2023). A tunable calcium phosphate coating to drive in vivo osseointegration of composite engineered tissues. Cells, tissues, organs, 212(5), 383-398.

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