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Streptavidin alexa488

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Streptavidin-Alexa488 is a fluorescent conjugate composed of streptavidin and the Alexa Fluor 488 dye. Streptavidin is a tetrameric protein that binds to biotin with high affinity. The Alexa Fluor 488 dye is a green-fluorescent dye with excitation and emission maxima of 490 nm and 525 nm, respectively.

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Lab products found in correlation

Rabbit anti iba1, by Fujifilm (2 mentions) Rhodamine phalloidin, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Staurosporine, by Merck Group (1 mentions) Chloroform, by Avanti Polar Lipids (1 mentions) Af6034, by R&D Systems (1 mentions) Fluoview confocal microscope, by Olympus (1 mentions) Mouse anti rat mhc 2, by Bio-Rad (1 mentions) Accumax, by Innovative Cell Technologies (1 mentions) Accutase, by Innovative Cell Technologies (1 mentions) Rabbit anti s100β, by Abcam (1 mentions) Rat anti cd68, by BioLegend (1 mentions) Goat anti gfap, by Santa Cruz Biotechnology (1 mentions) Mouse anti biotin, by Jackson ImmunoResearch (1 mentions) Rabbit anti tgf β1, by Abcam (1 mentions) Goat anti iba1, by Abcam (1 mentions) Cy3 conjugated donkey anti rabbit antibody, by Jackson ImmunoResearch (1 mentions) Biotinylated anti mouse igg, by Vector Laboratories (1 mentions) Pv235, by Swant (1 mentions)

7 protocols using streptavidin alexa488

1

Annexin V and Lipid Membrane Binding

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The following reagents were used: AnnexinV-Alexa-568 (IF 1:50, Invitrogen; A13202), Streptavidin-Alexa-488 (IF 2 µg/mL in PBS, Jackson ImmunoResearch Laboratories; 016-540-084), Phalloidin Rhodamine, Alexa Fluor 488 or Alexa Fluor 647 (IF 1:200, Molecular Probes; R415, A12379, A22287, respectively), Staurosporine (Sigma-Aldrich; S6942). The following lipids were used in this study: POPC, DOPS and DSPE-PEG (2000) Biotin all stored in Chloroform (Avanti Polar Lipids; 850457, 840035 and 880129, respectively).
Elfmark L.A., Wenzel E.M., Wang L., Pedersen N.M., Stenmark H, & Raiborg C. (2023). Protrudin-mediated ER-endosome contact sites promote phagocytosis. Cellular and Molecular Life Sciences, 80(8), 216.
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2

Imaging Mesenteric Artery Structure with Nogo-B

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Following full relaxation with Ach (1 × 10−6 M), mesenteric arteries were fixed with 4% PFA and left overnight at 4 °C. The elastic lamina of paraffin-embedded mesenteric arteries was visualized with Verhoeff's Van Gieson (VVG) staining. Wall thickness, area and radius were determined by using a computerized image-analysis system (Image-Pro).
PFA-fixed mesenteric arteries were paraffin-embedded or OCT-embedded. For immunohistochemistry, following deparaffinization and antigen retrieval, mesenteric artery sections were incubated overnight with antibody against Nogo-B (#AF6034, R&D, 1:100) followed by anti-sheep antibody (#713-065-003, Jackson ImmunoResearch, 1:200). Staining was developed with diaminobenzidene (DAB). Finally, the sections were counterstained with Mayer's hematoxylin solution.
For immunofluorescence, frozen mesenteric artery sections were stained for Nogo-B (1:100, R&D) and biotinylated IB4 (1:100, BD Biosciences) overnight at 4 °C, and were then stained with Cy3-labeled anti-sheep antibody (#A21436, Invitrogen, 1:200) and streptavidin–Alexa 488 (#016-540-084, Jackson ImmunoResearch, 1:200) in PBS for 1 h. Nuclei were stained with DAPI. Confocal immunofluorescence images of the tissues were captured on an Olympus Fluoview confocal microscope.
Cantalupo A., Zhang Y., Kothiya M., Galvani S., Obinata H., Bucci M., Giordano F.J., Jiang X.C., Hla T, & Di Lorenzo A. (2015). Nogo-B regulates endothelial sphingolipid homeostasis to control vascular function and blood pressure. Nature medicine, 21(9), 1028-1037.
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3

Dual-Label Immunostaining of Dopamine 1 Receptor and MHC II in Rat Brain

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Free-floating, paraformaldehyde-fixed brain sections, were stained with rabbit anti rat dopamine 1 receptor (diluted to 1:200, Abcam, Cambridge, UK) and mouse anti rat MHC II (diluted to 1:100, AbD Serotec, Düsseldorf, Germany) at 4°C overnight after blocking in 5% normal serum for one hour at rt. Thereafter, sections were incubated with secondary swine anti rabbit biotinylated antibody (diluted to 1:400, Jackson ImmunoResearch, Newmarket, UK) for one hour at rt, followed by incubation with Streptavidin Alexa 488 (diluted to 1:400) and donkey anti mouse Cy3 conjugated (diluted to 1:400, Jackson ImmunoResearch, Newmarket, UK) for 90 minutes at rt.
Kuric E, & Ruscher K. (2014). Dynamics of major histocompatibility complex class II-positive cells in the postischemic brain - influence of levodopa treatment. Journal of Neuroinflammation, 11, 145.
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4

Purification of Primary Mammary Epithelial Cells

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Primary pMEC cultures were expanded for 13-14d post-isolation with two differential trypsinizations using Accutase (Innovative Cell Technologies, Mira Mesa, CA) to remove fibroblasts, followed by Accumax (Innovative Cell Technologies) to dislodge pMEC. Single cells were incubated with phycoerythrin-conjugated anti- CD140a (BD Biosciences, Franklin Lakes, NJ) and/or biotinylated anti-human CD49f (AbD Serotec, Oxford, UK), followed by streptavidin-Alexa 488 (Jackson Immunoresearch) and propidium iodide (10 μg/ml) and sorted using a MoFlo cell sorter (Cytomation, West Lafayette, IN).
Rowson-Hodel A.R., Manjarin R., Trott J.F., Cardiff R.D., Borowsky A.D, & Hovey R.C. (2015). Neoplastic transformation of porcine mammary epithelial cells in vitro and tumor formation in vivo. BMC Cancer, 15, 562.
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5

Immunohistochemistry of Neurodegenerative Markers

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Serial sections (1:24) were antigen retrieved (10 mM sodium citrate, 80 °C, 30 min), washed (1X phosphate buffered saline), blocked (5% Donkey Serum with 0.3% Triton-X), and incubated for 3 days at 4 °C with primary antibodies. Primary antibodies include rabbit anti-IBA1 (Wako, cat: 016-20001; 1:500), goat anti-IBA1 (Abcam, cat: ab107159; 1:1500), goat anti-GFAP (Santacruz Biotech, cat: sc-6170; 1:250), goat anti-GFAP (Novus Biological, cat: NB100-53809; 1:2000), rabbit anti-TGFβ1 (Abcam, cat: ab215715; 1:250), rat anti-CD68 (Biolegend, cat: 137002, 1:400), goat anti-Nestin (Novus biologicals, cat: NB100-1604, 1:400), rabbit anti-S100β (Abcam, cat: ab41548; 1:1500) and mouse anti-amyloid beta (6F3D) (Dako, cat: M0872; 1:200). After washing, antibodies were incubated in secondary antibodies (Jackson Immunoresearch; 1:200) for 1 h at room temperature, washed, and mounted. For plaque staining, sections were pre-treated for 5 min in 10% formic acid, followed by 10 min in 0.1 M borate buffer (PH 8.0). After 3 days of primary antibody, sections were incubated with anti-mouse biotin (Jackson ImmunoResearch, cat: 715-065-150; 1:200) for 2 h at room temperature, washed, and incubated with streptavidin-Alexa488 (Jackson ImmunoResearch, cat: 016-540-084; 1:400) for 1 h at room temperature, washed, and mounted.
Silburt J, & Aubert I. (2022). MORPHIOUS: an unsupervised machine learning workflow to detect the activation of microglia and astrocytes. Journal of Neuroinflammation, 19, 24.
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6

Characterization of Parvalbumin-Expressing GABAergic Neurons

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Transversal slices (350 μm) from somatosensory cortex were prepared (Kotzadimitriou et al., 2018 (link)) from 4- to 6-week-old heterozygous male CB6-Tg(Gad1-EGFP)G42Zjh/J -mice (The Jackson Laboratory, stock 007677, GAD67-GFP G42 line) expressing td-Tomato fluorophore preferably in parvalbumin GABAergic neurons (Chattopadhyaya et al., 2004 (link)). Cells were confirmed to be fast-spiking showing fast spike kinetics and a high-frequency non-accommodation firing pattern for suprathreshold depolarizing 500 ms pulses. Cells were visualized with streptavidin Alexa488 (1:2000, Jackson ImmunoResearch Lab, Inc) and analyzed by eye under epifluorescence microscopy to exclude axo-axonic cells. Three cells were selected for pv immunoreactivity, and they were all immunopositive for pv (see Supplementary file 1).
Szegedi V., Paizs M., Baka J., Barzó P., Molnár G., Tamas G, & Lamsa K. (2020). Robust perisomatic GABAergic self-innervation inhibits basket cells in the human and mouse supragranular neocortex. eLife, 9, e51691.
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7

Immunohistochemical Labeling of Parvalbumin and Microglia in Rat Brain

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Rats were perfused at the end of the fourth week with physiological saline, followed by a 4% paraformaldehyde solution. Details of the procedure are described elsewhere [6] . 80-m-thick coronal cryosections through the brain specimens were collected in 0.1 M phosphate buffer (pH 7.3). All sections along the rostrocaudal aspect of the parvafox nucleus (in general 28 [range: 25-30] sections, spanning a total length of 2.24 [range: 2.0-2.4] mm) were incubated first with a monoclonal primary antibody against PV (PV235, 1:1000, Swant, Marly, Switzerland) for 24-48 h at 4 • C, then with biotinylated anti-mouse IgG (1:200, Vector Laboratories, Burlingame, CA) for 2 h at ambient temperature, and finally with streptavidin-Alexa488 (1:200, Jackson Immunoresearch Laboratory, West Grove, PA) for 3 h at ambient temperature. In addition, every fourth section of the specimens from rats in cohort two was exposed first to rabbit anti-Iba1 (ionized calcium-binding adaptor molecule 1; Wako Pure Chemicals, Osaka, Japan [0.25 g/ml]) for 24 h at 4 • C and then to Cy3-conjugated donkey anti-rabbit antibody (Jackson Immunoresearch Laboratory [1:200] ) for 2 h at ambient temperature.
Roccaro-Waldmeyer D.M., Babalian A., Müller A, & Celio M.R. (2016). Reduction in 50-kHz call-numbers and suppression of tickling-associated positive affective behaviour after lesioning of the lateral hypothalamic parvafox nucleus in rats. Behavioural brain research, 298(Pt B).
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