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Donkey anti chicken 488

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Donkey anti-chicken 488 is a secondary antibody conjugated with the fluorescent dye Alexa Fluor 488. It is designed to detect and bind to chicken antibodies in laboratory experiments.

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Lab products found in correlation

Donkey anti mouse 647, by Jackson ImmunoResearch (4 mentions) Donkey anti rabbit 647, by Jackson ImmunoResearch (4 mentions) Lsm 880 confocal microscope, by Zeiss (4 mentions) Chicken anti gfp, by Abcam (3 mentions) Chicken anti gfp, by Thermo Fisher Scientific (3 mentions) Mouse anti pkc δ, by BD (2 mentions) Chicken anti gfp, by Aves Labs (2 mentions) Normal goat serum (ngs), by Jackson ImmunoResearch (2 mentions) Gold antifade reagent, by Thermo Fisher Scientific (2 mentions) Rat anti mcd8a, by Thermo Fisher Scientific (2 mentions) Mcd0800, by Abcam (2 mentions) Goat anti rat 488, by Thermo Fisher Scientific (2 mentions) Rabbit anti dsred, by Takara Bio (2 mentions) Vs120 slide scanner microscope, by Olympus (2 mentions) Rabbit anti pstat3, by Cell Signaling Technology (2 mentions) Donkey anti goat 647, by Thermo Fisher Scientific (1 mentions) Donkey anti goat cy3, by Jackson ImmunoResearch (1 mentions) Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions) Normal donkey serum (nds), by Jackson ImmunoResearch (1 mentions) I32450, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Alexa 488 donkey anti chicken antibody Alexa Fluor 488-conjugated donkey anti-chicken IgY Alexa Fluor 488 donkey anti-chicken secondary antibody AlexaFluor 488 donkey anti-chicken IgY Alexa Fluor 488-AffiniPure donkey anti-chicken IgY (IgG) (H+L), Alexa Fluor 488-donkey anti-chicken IgG Alexa 488-conjugated donkey anti-chicken secondary antibody Alexa Fluor 488 donkey anti-chicken IgY (IgG) (H+L) Alexa 488 donkey anti-chicken IgG Alexa Fluro 488 donkey anti-chicken

16 protocols using donkey anti chicken 488

1

Immunofluorescent Labeling of Neural Markers

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Primary antibodies used for immunofluorescence were PDGFRα (1:200, R&D Systems, AF1062), Olig2 (1:200, Millipore, MABN50), GFP-488 (1:400, Fisher Scientific, A21311), and GFP (1:1000, Invitrogen, A10262). Secondary antibodies used were Donkey anti-Goat Cy3 (2 μg/mL, Jackson ImmunoResearch, 705-165-147), Donkey anti-Mouse 647 (2 μg/mL, Jackson ImmunoResearch, 715-605-150), Donkey anti-Mouse 546 (2 μg/mL, Life Technologies, A10036), Donkey anti-Chicken 488 (2 μg/mL, Jackson ImmunoResearch, 703-545-155), and Donkey anti-Goat 647 (2 μg/mL, Invitrogen, A21447).
Beiter R.M., Rivet-Noor C., Merchak A.R., Bai R., Johanson D.M., Slogar E., Sol-Church K., Overall C.C, & Gaultier A. (2022). Evidence for oligodendrocyte progenitor cell heterogeneity in the adult mouse brain. Scientific Reports, 12, 12921.
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2

Immunohistochemical Analysis of Pnoc-IRES-Cre Mice

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For processing tissue samples for immunohistochemistry, mice were euthanized with pentobarbital (50 mg.kg, 1.p.) and transcardially perfused with 0.01 M phosphate-buffered saline (PBS) and 4% paraformaldehyde (PFA). Tissue was fixed overnight in PFA at 4°C, cryoprotected with 30% sucrose in PBS, and 40 μm thick coronal sections were collected with a cryostat. Immunochemistry was performed in Pnoc-IRES-Cre mice using the following primary (kept overnight at 4°C) and secondary (kept at room temperature for 2 h) antibodies: chicken-anti-GFP (1:1,000; Aves labs, Tigard, OR), donkey anti-chicken 488 (1:500; Jackson Immuno Research Labs, West Grove, PA), mouse anti-PKCδ (1:500; BD Biosciences, Fanklin Lakes, NJ), donkey anti-mouse 647 (1:500; Jackson Immuno Research Labs, West Grove, PA), rabbit anti-Somatostatin (1:2,000; BMA Biomedicals, Switzerland), and donkey anti-rabbit 647 (1:500; Jackson Immuno Research Labs, West Grove, PA). Antibodies used with dilutions can also be found in supplementary information (Table S1). Immunoprocessing procedures were done as previously described (Jennings et al., 2013a (link)), and sections were counterstained with DAPI and coverslipped for subsequent confocal microscopy and counted using ImageJ software.
Rodriguez-Romaguera J., Ung R.L., Nomura H., Otis J.M., Basiri M.L., Namboodiri V.M., Zhu X., Robinson J.E., van den Munkhof H.E., McHenry J.A., Eckman L.E., Kosyk O., Jhou T.C., Kash T.L., Bruchas M.R, & Stuber G.D. (2020). Prepronociceptin-Expressing Neurons in the Extended Amygdala Encode and Promote Rapid Arousal Responses to Motivationally Salient Stimuli. Cell reports, 33(6), 108362.
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3

Immunohistochemistry of Drosophila Brains

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Fly brains were dissected in 1X PBS and fixed for 20 min at room temperature in 1X PBS containing 4% formaldehyde (Sigma: F8775). Fixed brains were washed 3 X 20 min in 1X PBS with 0.5% Triton X-100 (PBST; Sigma: T8787), followed by incubation in PBST with 5% normal goat serum (blocking solution; Jackson ImmunoResearch: 005-000-121) for 30 min. Brains were then incubated overnight with primary antibodies in blocking solution at 4°C, washed 3 X 20 min in PBST, before incubating overnight with secondary antibodies in PBST at 4°C. Brains were then washed 3 X 20 min in PBST and mounted on glass slides in Gold anti-fade reagent (Thermo Fisher Scientific: S36937). Antibodies used in this study are Mouse anti-Brp (1:50; DSHB: nc82), Rat anti-mCD8a (1:100; Thermo Fisher Scientific: MCD0800, lot# 1968949), chicken anti-GFP (1:5000; Abcam: 13970, lot# GR23665112), rabbit anti-Dsred (1:500; Clontech: 632496, lot# 1509043), donkey anti-chicken-488 (1:400; Jackson ImmunoResearch: 131753, lot# 131753), Goat anti-rat-488 (1:400; Thermo Fisher Scientific: A11006, lot# 1728142) and Goat anti-Rabbit-Cy3 (1:400; Jakson ImmumoResearch: 111-165-144, lot# 123834). Brains were imaged using an LSM 880 confocal microscope (Zeiss) and images were analyzed in Fiji/ImageJ61 (link).
Senapati B., Tsao C.H., Juan Y.A., Chiu T.H., Wu C.L., Waddell S, & Lin S. (2019). A neural mechanism for deprivation state-specific expression of relevant memories in Drosophila. Nature neuroscience, 22(12), 2029-2039.
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4

Leptin-Induced pSTAT3 Activation Imaging

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Mice were injected with 5 mg/kg recombinant leptin two hours prior to perfusion (as above). Brain sections were washed in 0.1 M phosphate-buffered saline pH 7.4 followed by incubation in 5% NaOH and 0.3% H2O2 for 2 min, then with 0.3% glycine (10 min), and finally with 0.03% SDS (10 min), all made up in PBS. Sections were blocked in 3% normal donkey serum/0.25% Triton X-100 in PBS for 1 hour at room temperature and then incubated overnight at room temperature in blocking solution containing 1/250 rabbit anti-pSTAT3 (Cell Signalling, #9145) and 1/1000 chicken anti-GFP (Life Technologies, #A10262). The next morning sections were extensively washed in PBS and then incubated in 1/250 donkey anti-rabbit 594 (Molecular Probes, R37119) and 1/1000 donkey anti-chicken 488 (Jackson ImmunoResearch, 703-545-155) for 2 h at room temperature. After several washes in PBS, sections were mounted onto gelatin-coated slides and fluorescent images were captured with Olympus VS120 slide scanner microscope.
Garfield A.S., Shah B.P., Burgess C.R., Li M.M., Li C., Steger J.S., Madara J.C., Campbell J.N., Kroeger D., Scammell T.E., Tannous B.A., Myers MG J.r., Andermann M.L., Krashes M.J, & Lowell B.B. (2016). Dynamic GABAergic afferent modulation of AgRP neurons. Nature neuroscience, 19(12), 1628-1635.
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5

Leptin-Induced pSTAT3 Activation Imaging

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Mice were injected with 5 mg/kg recombinant leptin two hours prior to perfusion (as above). Brain sections were washed in 0.1 M phosphate-buffered saline pH 7.4 followed by incubation in 5% NaOH and 0.3% H2O2 for 2 min, then with 0.3% glycine (10 min), and finally with 0.03% SDS (10 min), all made up in PBS. Sections were blocked in 3% normal donkey serum/0.25% Triton X-100 in PBS for 1 hour at room temperature and then incubated overnight at room temperature in blocking solution containing 1/250 rabbit anti-pSTAT3 (Cell Signalling, #9145) and 1/1000 chicken anti-GFP (Life Technologies, #A10262). The next morning sections were extensively washed in PBS and then incubated in 1/250 donkey anti-rabbit 594 (Molecular Probes, R37119) and 1/1000 donkey anti-chicken 488 (Jackson ImmunoResearch, 703-545-155) for 2 h at room temperature. After several washes in PBS, sections were mounted onto gelatin-coated slides and fluorescent images were captured with Olympus VS120 slide scanner microscope.
Garfield A.S., Shah B.P., Burgess C.R., Li M.M., Li C., Steger J.S., Madara J.C., Campbell J.N., Kroeger D., Scammell T.E., Tannous B.A., Myers MG J.r., Andermann M.L., Krashes M.J, & Lowell B.B. (2016). Dynamic GABAergic afferent modulation of AgRP neurons. Nature neuroscience, 19(12), 1628-1635.
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6

Retinal Tissue Labeling and Imaging

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Eyeballs of P7 or P8 mice were fixed in 4% formaldehyde at 4 °C during 2 h and then washed in PBS and dissected. Retinas were permeabilized and blocked at 4 °C overnight in PBS, 1% BSA, 0.5% Triton X-100, 5% Normal donkey serum (Jackson ImmunoResearch). Ai3 and Ai14 retinas were incubated with isolectin B4 conjugated with Alexa Fluor-647 (IB4, ThermoFisher Scientific I32450, 1:200) in PBlec: PBS, 0,1 mM CaCl2, 0,1 mM MgCl2, 0,1 mM MnCl2, 1% Triton X-100 pH 6.8 for 4 h at room temperature. R26R-EYFP and mTmG retinas were also incubated with chicken anti-GFP (Abcam ab13970, 1:500) during that time. This was followed by three 30′ washes in PBS, 0.5% BSA, 0.25% Triton X-100. R26R-EYFP and mTmG retinas were then incubated overnight at 4 °C with the Donkey anti-chicken 488 (Jackson ImmunoResearch, 703-545-155) in PBS, 0.5% BSA, 0,25% Triton X-100 and washed again three times 30′ in PBS. Retinas were mounted with ProLong Gold Antifade Mountant (Life Technologies, P36930) and imaged on a confocal laser-scanning microscope (Leica TCS SP8) with a 10X magnification objective.
Álvarez-Aznar A., Martínez-Corral I., Daubel N., Betsholtz C., Mäkinen T, & Gaengel K. (2019). Tamoxifen-independent recombination of reporter genes limits lineage tracing and mosaic analysis using CreERT2 lines. Transgenic Research, 29(1), 53-68.
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7

Histology and Immunofluorescence of Brain Slices

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Perfusion and histology were performed as described in Garfield et al.153 (link). Brain slices (60 μm thick) were collected and one of every three consecutive slices was scanned. The primary antibodies used in this paper are chicken anti-GFP (1:1000, Invitrogen; used for GFP and GCaMP6s) and rat anti-mCherry (1:1000, ThermoFisher; used for mCherry and tdTomato). The secondary antibodies are Donkey anti-chicken 488 (1:1000, Jackson) and Donkey anti-rat 594 (1:1000, Jackson).
Zhang S.X., Kim A., Madara J.C., Zhu P.K., Christenson L.F., Lutas A., Kalugin P.N., Jin Y., Pal A., Tian L., Lowell B.B, & Andermann M.L. (2023). Competition between stochastic neuropeptide signals calibrates the rate of satiation. Research Square.
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8

Immunohistochemistry Staining Protocol

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Immunohistochemistry was performed as previously described (Sulkowski et al., 2011 (link)). Primary antibodies used were rabbit anti-PP2CB (used at 1:50 dilution) (Biorbyt); chicken anti-GFP (used at 1:1000 dilution) (Abcam); mouse anti-Cut (used at 1:100 dilution) (DSHB); mouse anti-Futsch (22C10) (used at 1:100 dilution) (Developmental Studies Hybridoma Bank); mouse anti-acetylated α-tubulin (used at 1:100) (Santa Cruz); rabbit anti-pS172 β-tubulin (used at 1:100 dilution) (ab78286, Abcam); rabbit anti-FoxO (used at 1:100 dilution) (ab195977, Abcam). Donkey anti-chicken 488 (1:1000) (Jackson Immunoresearch), donkey anti-rabbit 555 (1:200) (Life Technologies), donkey anti-mouse (1:200) (Life Technologies), and Alexa-Fluor goat anti-horseradish peroxidase (HRP) 647 (1:200) were used as secondary antibodies.
Bhattacharjee S., Lottes E.N., Nanda S., Golshir A., Patel A.A., Ascoli G.A, & Cox D.N. (2022). PP2A phosphatase regulates cell-type specific cytoskeletal organization to drive dendrite diversity. Frontiers in Molecular Neuroscience, 15, 926567.
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9

Immunostaining of Cholinergic and Glutamatergic Neurons

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Retinas were fixed and counterstained with the following antibodies. Primary antibodies: Goat anti-ChAT (Choline acetyltransferase; 1:200, Millipore Sigma #AB144); Rabbit anti-VGluT3 (1:250, Invitrogen #PA5-85784). Chicken anti-GFP (1:1000, Abcam #ab13970) was used to enhance the fluorescence of the Cre-dependent GFP virus. Rabbit anti-HA tag (1: 200, Cell Signaling Technology #3724) was used to stain the HA-tagged hM4Di receptor in the VGluT3 x DREADD mouse. Secondary antibodies: Donkey anti-Chicken 488 (1:1000, Jackson Immunoresearch #703-545-155); Donkey anti-Chicken 594 (1:1000, Jackson Immunoresearch #703-585-155); Donkey anti-Goat 488 (1:200, Invitrogen #A-11055); Donkey anti-Goat 594 (1:200, Invitrogen #A-11058); Donkey anti-Rabbit 647 (1:200, Invitrogen #A31573).
Mani A., Yang X., Zhao T.A., Leyrer M.L., Schreck D, & Berson D.M. (2023). A circuit suppressing retinal drive to the optokinetic system during fast image motion. Nature Communications, 14, 5142.
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10

Immunohistochemical Analysis of Pnoc-IRES-Cre Mice

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For processing tissue samples for immunohistochemistry, mice were euthanized with pentobarbital (50 mg.kg, 1.p.) and transcardially perfused with 0.01 M phosphate-buffered saline (PBS) and 4% paraformaldehyde (PFA). Tissue was fixed overnight in PFA at 4°C, cryoprotected with 30% sucrose in PBS, and 40 μm thick coronal sections were collected with a cryostat. Immunochemistry was performed in Pnoc-IRES-Cre mice using the following primary (kept overnight at 4°C) and secondary (kept at room temperature for 2 h) antibodies: chicken-anti-GFP (1:1,000; Aves labs, Tigard, OR), donkey anti-chicken 488 (1:500; Jackson Immuno Research Labs, West Grove, PA), mouse anti-PKCδ (1:500; BD Biosciences, Fanklin Lakes, NJ), donkey anti-mouse 647 (1:500; Jackson Immuno Research Labs, West Grove, PA), rabbit anti-Somatostatin (1:2,000; BMA Biomedicals, Switzerland), and donkey anti-rabbit 647 (1:500; Jackson Immuno Research Labs, West Grove, PA). Antibodies used with dilutions can also be found in supplementary information (Table S1). Immunoprocessing procedures were done as previously described (Jennings et al., 2013a (link)), and sections were counterstained with DAPI and coverslipped for subsequent confocal microscopy and counted using ImageJ software.
Rodriguez-Romaguera J., Ung R.L., Nomura H., Otis J.M., Basiri M.L., Namboodiri V.M., Zhu X., Robinson J.E., van den Munkhof H.E., McHenry J.A., Eckman L.E., Kosyk O., Jhou T.C., Kash T.L., Bruchas M.R, & Stuber G.D. (2020). Prepronociceptin-Expressing Neurons in the Extended Amygdala Encode and Promote Rapid Arousal Responses to Motivationally Salient Stimuli. Cell reports, 33(6), 108362.
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