Sc 16128 r

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Sc-16128-R is a lab equipment product from Santa Cruz Biotechnology. It is a general-purpose laboratory instrument used for various scientific applications.

Automatically generated - may contain errors

Lab products found in correlation

Tcs sp5 aobs microscope, by Leica (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Alexa fluor 594, by Thermo Fisher Scientific (2 mentions) Axioplan, by Zeiss (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Ab6994, by Abcam (1 mentions) Ab4648, by Abcam (1 mentions) Ab177487, by Abcam (1 mentions) Ab6326, by Abcam (1 mentions) Ab7349, by Abcam (1 mentions) Mca1957ga, by Bio-Rad (1 mentions) Sc 393581, by Santa Cruz Biotechnology (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Ab5103, by Abcam (1 mentions) P36935, by Thermo Fisher Scientific (1 mentions) Dmi6000b, by Leica (1 mentions) Poly d lysine, by Merck Group (1 mentions) Photometric enzyme immunoassay, by Roche (1 mentions) Flexstation 3 microplate reader, by Molecular Devices (1 mentions) Quant it picogreen kit, by Thermo Fisher Scientific (1 mentions)

6 protocols using sc 16128 r

Extracellular Traps in Kidney Tissue

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Samples of kidney tissue were collected from mice, embedded in Tissue-Tek® O.C.T.™, immediately frozen in 2-methylbutane (isopentane) resting in liquid nitrogen and stored at −70°C. Sections were cut to a thickness of 10–15 μm, fixed with 4% paraformaldehyde and stained with Hoechst 33342 (1 μg/mL, Molecular Probes), anti-MPO (1:50; sc-16128-R, Santa Cruz Biotechnology) and anti-DNA/histone H1 (1:200, MAB3864, Merck Millipore) antibodies, followed by anti-rabbit Alexa Fluor 594 (1:100; Molecular Probes) or anti-mouse Alexa Fluor 488 (1:400; Molecular Probes). Confocal images were taken with a Leica TCS SP5-AOBS microscope (Leica Microsystems, Mannheim, Germany); epifluorescence images were taken with a Zeiss Axioplan. Permeabilizing agents were not used, resulting in exclusive extracellular labeling. Assessment was made in a purely qualitative fashion to detect the presence or absence of NETs in the analyzed samples.

Czaikoski P.G., Mota J.M., Nascimento D.C., Sônego F., Castanheira F.V., Melo P.H., Scortegagna G.T., Silva R.L., Barroso-Sousa R., Souto F.O., Pazin-Filho A., Figueiredo F., Alves-Filho J.C, & Cunha F.Q. (2016). Neutrophil Extracellular Traps Induce Organ Damage during Experimental and Clinical Sepsis. PLoS ONE, 11(2), e0148142.

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Immunohistochemical Analysis of Brain Markers

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Samples were blocked with a solution containing 0.3% Triton, 3% goat serum, and 1% bovine serum albumin. To label BrdU, sections were preincubated with 2N HCL at 37 °C, followed by incubation with rat monoclonal antibody against BrdU (1:400; ab6326; Abcam, UK). For others, sections were incubated with antibody against TREM-1 (1:200; ab217161, Abcam, UK), Iba-1 (1:500; #019-19741, Wako, Japan), CD68 (1:200; MCA1957GA, AbD Serotec, UK), GFAP (1:500; ab4648, Abcam, UK), NeuN (1:500; ab177487, Abcam, UK), MBP (1:500; ab7349, Abcam, UK), vWF (1:500; ab6994, Abcam, UK), MPO (1:200; sc-16128-R, Santa Cruz Biotechnology, USA), SYK (1:200; #13198, Cell Signaling Technology, USA), and Gasdermin D (GSDMD) (1:200; sc-393581, Santa Cruz Biotechnology, UK) overnight at 4 °C. Sections were then incubated with appropriate secondary antibodies and DAPI (Sigma-Aldrich, USA). The positive signal was quantified by Image J software (NIH, USA).

Xu P., Zhang X., Liu Q., Xie Y., Shi X., Chen J., Li Y., Guo H., Sun R., Hong Y., Liu X, & Xu G. (2019). Microglial TREM-1 receptor mediates neuroinflammatory injury via interaction with SYK in experimental ischemic stroke. Cell Death & Disease, 10(8), 555.

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Immunofluorescence Analysis of Neutrophil Extracellular Traps

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The tissue segment was embedded in optimum cutting temperature, and sections (15 μm) were blocked and incubated with primary antibodies against Histone H3 citrulline R17+R2+R8 (1:1000; ab5103, Abcam) and then stained with conjugated secondary antibodies. Images and analysis were performed using a fluorescence microscope (Leica DMI6000B). For immunostaining, neutrophils were attached on slides coated with poly-d-lysine (Sigma) and stimulated with PMA (100 ng/ml) or medium. The slides were then fixed with 4% paraformaldehyde and stained with DAPI (P36935, Molecular Probes), anti-MPO (1:50; sc-16128-R, Santa Cruz Biotechnology), and anti-histone H4 (1:200, sc-25260, Santa Cruz Biotechnology) antibodies, followed by anti-rabbit Alexa Fluor 594 (1:100; Molecular Probes) or anti-mouse Alexa Fluor 488 (1:400; Molecular Probes). Confocal images were taken with a Leica TCS SP5-AOBS microscope (Leica Microsystems, Mannheim, Germany). Epifluorescence images were taken with a Zeiss Axioplan. Permeabilizing agents were not used for exclusive extracellular labeling. The percentage of NETs was determined from six non-overlapping fields per well and the average was from triplicates for each condition in every experiment. All analysis was performed blinded to treatment conditions.

Colón D.F., Wanderley C.W., Franchin M., Silva C.M., Hiroki C.H., Castanheira F.V., Donate P.B., Lopes A.H., Volpon L.C., Kavaguti S.K., Borges V.F., Speck-Hernandez C.A., Ramalho F., Carlotti A.P., Carmona F., Alves-Filho J.C., Liew F.Y, & Cunha F.Q. (2019). Neutrophil extracellular traps (NETs) exacerbate severity of infant sepsis. Critical Care, 23, 113.

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Quantification of Histone-DNA and MPO-DNA Complexes in Serum

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The levels of human histone (H1, H2A, H2B, H3, and H4)-associated DNA fragments in the serum were quantified by a photometric enzyme immunoassay (Roche Life Science, IN, USA); and analyzed as determined by the manufacturer.
For the quantification of the DNA-MPO complex levels in the serum samples, a previously described ELISA was performed [19 (link)] The anti-MPO antibody used in our assays was a different one (catalog number sc-16128-R; Santa Cruz Biotechnology, TX, USA).

da Silva C.O., Dias A.A., da Costa Nery J.A., de Miranda Machado A., Ferreira H., Rodrigues T.F., Sousa Santos J.P., Nadaes N.R., Sarno E.N., Saraiva E.M., Schmitz V, & Pessolani M.C. (2019). Neutrophil extracellular traps contribute to the pathogenesis of leprosy type 2 reactions. PLoS Neglected Tropical Diseases, 13(9), e0007368.

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Quantifying DNA Binding to Myeloperoxidase

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This procedure was performed as previously described [41 (link)]. Briefly, an antibody bound to the 96-well clear-bottom black plate captured the enzyme MPO (5 μg/ml; sc-16128-R, Santa Cruz Biotechnology), and the amount of DNA bound to the enzyme was quantified using the Quant-iT™ PicoGreen® kit (Invitrogen) according to the manufacturer’s instructions. The fluorescence intensity (excitation at 488 nm and emission at 525 nm wavelength) was quantified by a FlexStation 3 Microplate Reader (Molecular Devices, CA, USA).

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Protein Analysis by SDS-PAGE and Western Blot

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Protein analysis was performed by SDS-PAGE electrophoresis/western blot, loading 30 μg each sample. Proteins were transferred to a nitrocellulose membrane (Amersham, GE Healthcare Europe GmbH, Milano, Italy), and revealed by immunoblotting with specific antibodies: rabbit polyclonal heme oxygenase-1 (HO-1) (1:200)(sc-10789; Santa Cruz Biotechnology™, Dallas, TX, USA), rabbit polyclonal inducible nitric oxide synthase (iNOS) (1:200) (sc-8310; Santa Cruz Biotechnology™), rabbit polyclonal cytochrome 1b1 (Cyp1b1) (1:200) (sc-32882; Santa Cruz Biotechnology™), goat polyclonal heat shock protein 70 (Hsp70) (1:200)(sc-10-70; Santa Cruz Biotechnology™), rabbit polyclonal cyclooxygenase 2 (COX2) (1:1000)(#4842; Cell Signalling Technology^®, Danvers, MA, USA), and rabbit polyclonal myeloperoxidase (MPO) (1:200) (sc-16128-R; Santa Cruz Biotechnology™). The secondary antibodies were appropriate horseradish peroxidase (HRP)-conjugated goat anti-rabbit (1:5000) (31460 Thermofisher Scientific™ Waltham, MA, USA) or donkey anti-goat (1:2000) (sc-2020; Santa Cruz Biotechnology™). Immunoblot bands have been analyzed and the optical density quantified by LAS4000 (GE Healthcare, Marlborough, MA, USA); all the data have been normalized to Ponceau staining (Sigma Chemical Co., Milano, Italy) [78 (link)].

Farina F., Lonati E., Milani C., Massimino L., Ballarini E., Donzelli E., Crippa L., Marmiroli P., Botto L., Corsetto P.A., Sancini G., Bulbarelli A, & Palestini P. (2019). In Vivo Comparative Study on Acute and Sub-acute Biological Effects Induced by Ultrafine Particles of Different Anthropogenic Sources in BALB/c Mice. International Journal of Molecular Sciences, 20(11), 2805.

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