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Appropriate fluorescence conjugated secondary antibodies

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Appropriate fluorescence conjugated secondary antibodies are laboratory reagents designed to detect and visualize specific target proteins or molecules in various immunoassay and imaging applications. These antibodies are chemically conjugated with fluorescent dyes, enabling the detection and localization of target analytes through fluorescence microscopy or flow cytometry techniques.

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Lab products found in correlation

Ox 42, by Abcam (2 mentions) Mannose receptor cd206, by Abcam (1 mentions) Glial fibrillary acidic protein (gfap), by Abcam (1 mentions) Neuronal nuclei (neun), by Abcam (1 mentions) Leica dmi8, by Leica Microsystems (1 mentions) Mouse anti neun, by Abcam (1 mentions) Rabbit anti ntn 1, by Abcam (1 mentions) Leica dmi8, by Leica camera (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions) Mannose receptor, by Abcam (1 mentions) Magnafire sp 2.1b software, by Olympus (1 mentions) Bx51 microscope, by Olympus (1 mentions) Anti icam 1, by Santa Cruz Biotechnology (1 mentions) Anti smemb, by Abcam (1 mentions) Magnafire sp 2.1b, by Olympus (1 mentions) Anti p p38 mapk, by Santa Cruz Biotechnology (1 mentions) Anti pdgfr β antibody, by Santa Cruz Biotechnology (1 mentions) Anti smooth muscle actin, by Santa Cruz Biotechnology (1 mentions) Cm3050, by Leica Microsystems (1 mentions)

4 protocols using appropriate fluorescence conjugated secondary antibodies

1

Immunohistochemical Analysis of Microglia

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Tissues were embedded in Optimal Cutting Temperature Compound (OCT, Fisher Scientific) and then sectioned into 10 μm slices. Sections were stained with OX-42 (1:1000, Abcam), mannose receptor (CD206) (1:500, Abcam), GFAP (1:1000, Abcam), and NeuN (1:1000, Abcam) at 4°C overnight. Appropriate fluorescence conjugated secondary antibodies (Jackson Immunoresearch, West Grove, PA) were incubated for 1 h at room temperature. Leica DMi8 (Leica Microsystems, Germany) was used to image the peri-ventricular region. M2 microglia activated cells were counted on 6 sections in every group over a 20 × microscopic field and measured as cells/field, as described 31 (link).
Zhang Y., Ding Y., Lu T., Zhang Y., Xu N., Yu L., McBride D.W., Flores J.J., Tang J, & Zhang J.H. (2017). Bliverdin Reductase-A Improves Neurological Function in a Germinal Matrix Hemorrhage Rat Model. Neurobiology of disease, 110, 122-132.
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2

Dual Immunofluorescence Staining of NTN-1 and NeuN

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Double fluorescence staining was performed as described previously (Huang et al., 2015 (link)). Frozen coronal sections (10 μm) were blocked with 5% donkey serum for 1 h and incubated at 4°C overnight with primary antibodies: rabbit anti-NTN-1 (1:500, Abcam), and mouse anti-NeuN (1:500, Abcam) followed by incubation with appropriate fluorescence-conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA) for 2 h at room temperature. Negative control staining was conducted by omitting the primary antibody. Finally, slides were covered with DAPI (Vector Laboratories, Inc.). The sections were visualized under a fluorescence microscope Leica DMi8. Microphotographs were analyzed with LASX software.
Xie Z., Huang L., Enkhjargal B., Reis C., Wan W., Tang J., Cheng Y, & Zhang J.H. (2017). Intranasal administration of recombinant Netrin-1 attenuates neuronal apoptosis by activating DCC/APPL-1/AKT signaling pathway after subarachnoid hemorrhage in rats. Neuropharmacology, 119, 123-133.
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3

Microglial response in intracerebral hemorrhage

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10 µm thick slices were first stained with OX-42 (1:1000, ABcam) and mannose receptor (1:1000, ABcam) overnight at 4 °C, followed by incubation with appropriate fluorescence conjugated secondary antibodies (Jackson Immunoresearch, West Grove, PA). The peri-hemorrhagic area was imaged by a Fluorescent Olympus-BX51 microscope and analyzed using MagnaFire SP 2.1B software (Olympus, Melville, NY). At least six sections per animal group over a microscopic field of 20 × (for microglia) were averaged and expressed as cells/field, as described (Wang and Dore, 2007 (link))
Flores J.J., Klebe D., Rolland W.B., Lekic T., Krafft P.R, & Zhang J.H. (2015). PPARγ-induced Upregulation of CD36 Enhances Hematoma Resolution and Attenuates Long-term Neurological Deficits after Germinal Matrix Hemorrhage in Neonatal Rats. Neurobiology of disease, 87, 124-133.
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4

Immunofluorescence Analysis of Cerebral Ischemia

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At 24 hours following ICH, mice were perfused under deep anesthesia with cold phosphate-buffered saline (PBS, pH 7.4), then infused with 10% formalin. Brains were removed and fixed in formalin at 4°C for a minimum of 3 days. Samples were dehydrated with 30% sucrose in phosphate-buffered saline (PBS, pH 7.4) and the frozen coronal slices (10 μm thick) were sectioned in cryostat (CM3050S; Leica Microsystems). Immunofluorescence was performed as previously described.22 (link),36 (link) Anti-PDGFR-β antibody (1:100, Santa Cruz), anti-p-p38 MAPK (1:100, Santa Cruz), anti-p-MK2 (1:100, Santa Cruz) anti-ICAM-1(1:100, Santa Cruz), anti-SMemb (1:100, abcam) were incubated separately with primary antibodies: anti-smooth muscle actin (1:1000, Santa Cruz) overnight at 4°C. It was then incubated with the appropriate fluorescence conjugated secondary antibodies (Jackson Immunoresearch, West Grove, PA). The slices were observed underneath a fluorescence microscope (Olympus BX51, Olympus Optical Co. Ltd, Japan), and pictures were taken with software MagnaFire SP 2.1B (Olympus, Melville, NY).
Yang P., Wu J., Miao L., Manaenko A., Matei N., Zhang Y., Xu L., Pearce W.J., Hartman R.E., Obenaus A., Zhang J.H., Xu F, & Tang J. (2016). Platelet-derived growth factor receptor-β regulates vascular smooth muscle cell phenotypic transformation and neuro-inflammation after intracerebral hemorrhage in mice. Critical care medicine, 44(6), e390-e402.
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