Brain or spinal cord sections were washed in PBS with Tween-20, pH 7.4 (PBST) and blocked in 3% normal donkey serum in PBST for 1 h at room temperature. Sections were then incubated overnight at room temperature in blocking solution containing primary antiserum (rat anti-mCherry, Life Technologies M11217, 1:1,000; rabbit anti-Pomc precursor, Phoenix Pharmaceuticals H-029–30, 1:1,000). The next morning, sections were extensively washed in PBS and then incubated in Alexa-fluorophore secondary antibody (Molecular Probes, A-21206, A-21209, all 1:1,000) for 1 h at room temperature. After several washes in PBS, brain sections were mounted on gelatin-coated slides. Spinal cord sections were counterstained and coverslipped with DAPI containing mounting medium (VECTASHIELD® Antifade Mounting Medium with DAPI, Cat# H-1200, Vector Laboratories).
Alexa fluorophore secondary antibody
Manufactured by Thermo Fisher Scientific
Alexa fluorophore secondary antibodies are fluorescently-labeled antibodies designed for use in immunoassays and other applications that require signal detection. They are conjugated with Alexa Fluor dyes, which provide bright, photostable fluorescence for sensitive detection of target proteins or molecules.
Automatically generated - may contain errors
Lab products found in correlation
Chicken anti gfp, by Thermo Fisher Scientific (7 mentions)
Vs120 slide scanner microscope, by Olympus (6 mentions)
Rabbit anti dsred, by Takara Bio (4 mentions)
Rat anti mcherry, by Thermo Fisher Scientific (3 mentions)
Sheep anti melanocyte stimulating hormone, by Merck Group (2 mentions)
Goat anti agrp, by Neuromics (2 mentions)
M11217, by Phoenix Pharmaceuticals (2 mentions)
H 029 30, by Neuromics (2 mentions)
Gt15023, by Thermo Fisher Scientific (2 mentions)
Bz x810 slide scanner microscope, by Keyence (2 mentions)
Zen blue software, by Zeiss (2 mentions)
H 029 30, by Phoenix Pharmaceuticals (1 mentions)
Vectashield antifade mounting medium with dapi, by Vector Laboratories (1 mentions)
Vs120slide scanner microscope, by Zeiss (1 mentions)
Observer z1 confocal microscope, by Zeiss (1 mentions)
Lsm 880 airyscan confocal microscope, by Zeiss (1 mentions)
Click it cell reaction buffer kit, by Thermo Fisher Scientific (1 mentions)
11 protocols using alexa fluorophore secondary antibody
1
Fluorescent Immunohistochemistry of Brain and Spinal Cord
2
Immunohistochemical Analysis of Neurotransmitter Systems
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Brain sections were washed in 0.1 M phosphate-buffered saline with Tween-20, pH 7.4 (PBST), 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 primary antiserum (sheep anti-melanocyte-stimulating hormone, Millipore (#5087) 1:1000; rabbit anti-dsRed, Clonetech (#632496)1:1000; chicken anti-GFP, Life Technologies (#A10262) 1:1,000; rabbit anti-oxytocin, Peninsula Laboratories (#T4084), 1/1000; goat anti-AgRP, Neuromics (#GT15023) 1/1000 or rabbit anti-FOS, Calbiochem (#PC38) 1/25,000). The next morning sections were extensively washed in PBS and then incubated in Alexa fluorophore secondary antibody (Molecular Probes, 1:1000) 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.
3
Immunofluorescence Labeling of Brain Sections
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Brain sections were washed in PBS with Tween-20, pH 7.4 (PBST) and blocked in 3% normal donkey serum/PBST for 1 h at room temperature. Then, brain sections were incubated overnight at room temperature in blocking solution containing primary antiserum (rat anti-mCherry, Life Technologies M11217, 1:1,000; rabbit anti-POMC precursor, Phoenix Pharmaceuticals H-029-30, 1:3,000, goat anti-AgRP, Neuromics GT15023, 1:3,000, chicken anti-GFP, Life Technologies A10262, 1:1,000, goat anti-Fos, Calbiochem PC38, 1:1,000). The next morning sections were extensively washed in PBS and then incubated in Alexa-fluorophore secondary antibody (Molecular Probes, 1:1,000) for 1 h at room temperature. After several washes in PBS, sections were mounted on gelatin-coated slides and fluorescence images were captured with Olympus VS120 slide scanner microscope.
4
Immunofluorescence Staining of Brain Sections
5
Immunofluorescent Labeling of Brain Sections
6
Immunohistochemical Analysis of Neurotransmitter Systems
7
Immunofluorescence Labeling of Brain Sections
8
Immunofluorescence Staining of Brain Sections
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Brain sections were washed in 0.1 M phosphate-buffered saline pH 7.4, 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 primary antiserum (rabbit anti-dsRed, Clonetech (#632496) 1:1000; chicken anti-GFP, Life Technologies (#A10262). The next morning sections were extensively washed in PBS and then incubated in Alexa fluorophore secondary antibody (Molecular Probes, 1:1000) 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. All primary antibodies used are validated for species and application (1DegreeBio and Antibody Registry).
9
Immunofluorescent Labeling of Brain Sections
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Brain sections were washed in 0.1 M phosphate buffered saline pH 7.4, blocked in 3% normal donkey serum/0.25% Triton X100 in PBS for 1 h at room temperature and then incubated overnight at room temperature in blocking solution containing primary antiserum (rabbit anti-GFP, 488-conjugated, Novus #NB600–308, 1:5000; rabbit anti-TH, Thermo Fisher Scientific #OPA1–04050, 1:5000). The next morning, sections were extensively washed in PBS before being mounted onto polarized slides. For TH staining, sections were first washed in PBS/.05% Tween-20, then washed in PBS before being incubated in Alexa fluorophore secondary antibodies (donkey anti-rabbit, Thermo Fisher Scientific #A-21206, 1:1000) for 1 h at room temperature. After several washes in PBS, sections were mounted onto polarized slides and fluorescent images were captured with a Keyence BZ-X810 slide scanner microscope. All primary and secondary antibodies used are validated for species and application (1DegreeBio and Antibody Registry).
10
Immunohistochemistry for TH and GFP
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Brain sections were washed in 0.1 M phosphate buffered saline pH 7.4, blocked in 3% normal donkey serum/0.25% Triton X100 in PBS for 1 h at room temperature and then incubated overnight at room temperature in blocking solution containing primary antiserum (rabbit anti-GFP, 488-conjugated, Novus #NB600-308, 1:5000; rabbit anti-TH, Thermo Fisher Scientific #OPA1-04050, 1:5000). The next morning, sections were extensively washed in PBS before being mounted onto polarized slides. For TH staining, sections were first washed in PBS/.05% Tween-20, then washed in PBS before being incubated in Alexa fluorophore secondary antibodies (donkey anti-rabbit, Thermo Fisher Scientific #A-21206, 1:1000) for 1 h at room temperature. After several washes in PBS, sections were mounted onto polarized slides and fluorescent images were captured with a Keyence BZ-X810 slide scanner microscope. All primary and secondary antibodies used are validated for species and application (1DegreeBio and Antibody Registry).
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