Sc 18355

Manufactured by Santa Cruz Biotechnology

Sc-18355 is a lab equipment product offered by Santa Cruz Biotechnology. It is a device designed to perform specific laboratory functions. The core function of this product is to facilitate certain experimental or analytical procedures within a laboratory setting.

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5 protocols using sc 18355

Immunohistochemical Profiling of Neural and Immune Markers

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The method has been described previously (29 (link)). Primary antibodies were to the following proteins: PDGFRα (1:200, rat, 558774, BD Biosciences), PDGFRα (1:8000, rabbit, gift from W. Stallcup, Sanford Burnham Prebys), Olig2 (1:2000, rabbit, gift from C.D. Stiles, Harvard), CD31 (1:200, rat, 553370, BD Biosciences), CD31 (1:200, rabbit, ab28346, Abcam), RNF43 (1:200, rabbit, ab84125, Abcam), WIF1 (1:200, rabbit, ab71204, Abcam), Caspase-3 (1:200, rabbit, 9661, Cell signaling), Aqp4 (1:100, mouse, ab9512, Abcam), PLVAP (1:100, rat, 553849, BD Pharmingen), Claudin5 (1:200, mouse, 352588, ThermoFisher), Fibrinogen (1:200, mouse, ab58207, Abcam), Iba1 (1:500, rabbit, SAG4318, Wako), CD11c (1:500, rabbit, MBS767644, MyBioSource), Arg1 (1:100, goat, sc-18355, Santa Cruz), iNOS (1:200, rabbit, PA3030A, ThermoFisher), CD3 (1:100, rabbit, ab5690, Abcam), CD4 (1:200, rabbit, ab183685, Abcam), CD8 (1:100, rat, ab22378, Abcam), F4/80 (1:500, rat, ab6640, Abcam), Ki67 (1:200, rabbit, ab16667, Abcam), SMI32 (1:1000, mouse, NE1023, Millipore), NF200 (1:1000, rabbit, N4142, Sigma), Wif1 (1:1000, rabbit, ab155101, Abcam).

Niu J., Tsai H.H., Hoi K.K., Huang N., Yu G., Kim K., Baranzini S.E., Xiao L., Chan J.R, & Fancy S.P. (2019). Aberrant oligodendroglial-vascular interactions disrupt the Blood Brain Barrier triggering CNS inflammation. Nature neuroscience, 22(5), 709-718.

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Immunohistochemical Profiling of Neural and Immune Markers

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Protein Expression Analysis by Western Blot

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The protein concentration was determined by bicinchoninic acid protein assay. Equal amount of protein was used, and Western blotting was performed as previously described (66 (link)). Primary antibodies included anti-CD9 antibody (ab92726, Abcam, MA), anti-CD63 antibody (216130, Abcam, MA), anti-CD81 antibody [10037, Cell Signaling Technology (CST)], anti–annexin A1 antibody (3299, CST), anti-HSP90 antibody (4877, CST), anti–IL-6R antibody (ab83053, Abcam, MA), anti-Stat3 antibody and anti-p-Stat3 antibody (9139S and 9145S, CST), anti–IL-6 antibody (12912, CST), anti–nuclear factor κB antibody (8242S, CST), anti–TNF-α antibody (ab1793, Abcam, MA), anti–IL-1b antibody (NB600-633, Novus), anti-CD86 antibody (ab119857, Abcam, MA), anti-Arg1 antibody (sc-18355, Santa Cruz Biotechnology, CA), and anti–β-actin antibody (ab28696, Abcam, MA). The gray intensity of protein blots was measured using ImageJ software (NIH).

Zhou T., He C., Lai P., Yang Z., Liu Y., Xu H., Lin X., Ni B., Ju R., Yi W., Liang L., Pei D., Egwuagu C.E, & Liu X. (2022). miR-204–containing exosomes ameliorate GVHD-associated dry eye disease. Science Advances, 8(2), eabj9617.

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Spinal Cord Immunofluorescence Staining

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Mice were anesthetized with avertin and perfused with PBS and 4% paraformaldehyde (PFA). Spinal cords were dissected and post-fixed in 4% PFA overnight at 4 °C and equilibrated in 20% sucrose. 20 μm-thick sections were pretreated with Reveal Decloaker Solution (RV1000M, Biocare Medical) for antigen retrieval according to the manufacturer’s instructions. After blocking, sections were incubated with primary antibodies overnight at 4 °C and followed by appropriate secondary antibodies conjugated with Alexa fluorescence 488 or 594. The following primary antibodies were used: MBP (SMI-99P, Covance, 1:300), NeuF200 (N4142, Sigma, 1:250), CS56 (C8035, Sigma, 1:250), MMP-2 (AB191677, Sigma, 1:500), laminin (L9393, Sigma, 1:500), CAT301 (MAB5284, Millipore, 1:250), versican (AB1032, Millipore, 1:250), Iba1 (019-19741, WAKO, 1:250), GFAP (MAB360, Millipore, 1:250), Olig2 (AB9610, Millipore, 1:250), CC1 (OP80, Millipore, 1:250), iNOS (#610329, BD Biosciences, 1:250), Arginase-1 (sc-18355, Santa Cruz, 1:50), PDGFRα (Cell signaling, #3174), and NG2 (Millipore, AB5320). For each staining, at least three individual animals/group were examined and images were captured with a Leica DFC500 fluorescence microscope. Staining was quantified using Image J software (US National Institutes of Health, USA). Fluorescence intensity was calculated as percentages of the mean value of the naive controls.

Luo F., Tran A.P., Xin L., Sanapala C., Lang B.T., Silver J, & Yang Y. (2018). Modulation of proteoglycan receptor PTPσ enhances MMP-2 activity to promote recovery from multiple sclerosis. Nature Communications, 9, 4126.

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Comprehensive Immunofluorescence Antibody Panel

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The primary antibodies were as follows: Olig2 (rabbit, AB9610, 1:500, Millipore; mouse, MABN50, 1:500, Millipore), MBP (rabbit, AB980, 1:750, Millipore), Adenomatus Polyposis Coli (APC/CC1) (mouse, OP80, 1:500, Calbiochem), GFAP (mouse, G3893, 1:1500, Sigma), Iba1 (rabbit, W1 W019-19741, 1:500, Wako), Arg-1 (goat, sc-18355, 1:100, Santa-Cruz), PDGFRα (rat, 558774, 1:500, BD Pharmingen), DHT (guinea-pig, GP-DHT1, 1 :200, Synabs), Neurofilament H (NF-H), Non-phosphorylated Smi-32 (mouse, 801701, 1 :300, Biolegend), Ki67 (mouse, 550609, 1:150, BD Pharmingen), Aromatase (rabbit, Ab18995, 1:200, Abcam), Claudin-5 (mouse, 35-2500, 1 :700, Invitrogen), pSTAT3 (Tyr705) (rabbit, 9145, 1: 500, Cell Signaling). AR is a home-made antibody (guinea-pig, Aa 283–298/Aa 406–420 from mus musculus AR Accession NP_038504.1; Eurogentec). The secondary antibodies were: goat anti-rabbit cyanine 3 conjugated (111165003, 1/250, Jackson Immunoresearch); goat anti-mouse Alexa 488 (A11029, 1:250, Thermo Fisher Scientific), goat anti-rabbit Alexa 633 (A21070, 1:750, Thermo Fisher Scientific), goat anti-rat Alexa 633 (A21094, 1:750, Thermo Fisher Scientific), goat anti-rabbit Alexa 488 (A32731, 1:350, Thermo Fisher Scientific); goat anti-guinea pig cyanine 3 conjugated (106165003, 1/500, Jackson Immunoresearch); donkey anti-goat Alexa 488 (A11055, 1:500, Thermo Fisher Scientific).

Zahaf A., Kassoussi A., Hutteau-Hamel T., Mellouk A., Marie C., Zoupi L., Tsouki F., Mattern C., Bobé P., Schumacher M., Williams A., Parras C, & Traiffort E. (2023). Androgens show sex-dependent differences in myelination in immune and non-immune murine models of CNS demyelination. Nature Communications, 14, 1592.

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