For immuno-fluorescence labeling, selected sections of hippocampus from bregma −1.8 and LEnt from bregma −2.8, were washed in PBS for 10 mins and then permeabilized in 0.5ml PBS/0.25% Triton X-100 (PBST). Block tissues in blocking solution supplemented with 5% BSA and 5% normal donkey semm in PBST, 1.5-2 hours at RT. Dilute 1° antibodies in 5% BSA/PBST and incubate sections for overnight at 4°C. On the second day, wash the brain sections 3x in PBST, 15 min each. And then incubate the brain sections in 2° antibodies (1:700 for Dylight-/Alexa-conjugated antibodies made in donkey purchased from Thermo Fisher Scientific) in 5% BSA/PBST and incubate for 2 hours at RT. For DAPI nuclei stain, dilute DAPI (1:10,000) in PBST and incubate for 15 min. Wash 2x with PBST then 1x with PBS, 10 min each. Mount the brain sections onto microscope glass slides in Prolong gold anti-fade reagent. The primary antibodies used in this study are as follows: NeuN (chicken, Millipore, cat# ABN91), 1: 300; MC1 (mouse, provided by Peter Davies, Northwell), 1:100; TOMA2 (mouse, provided by Rakez Kayed, UTMB Galveston), 1:200 [26 (link)]; Images were captured by Carl Zeiss confocal LSM700.
Dylight alexa conjugated antibodies
Manufactured by Thermo Fisher Scientific
DyLight-/Alexa-conjugated antibodies are fluorescently labeled secondary antibodies used for detection and visualization in various immunoassay and microscopy applications. These antibodies are designed to bind to primary antibodies and emit specific wavelengths of light when excited, enabling the identification and localization of target proteins or cells.
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Lab products found in correlation
3 protocols using dylight alexa conjugated antibodies
1
Hippocampal and Enthorhinal Cortex Immunofluorescence
2
Immunofluorescence Labeling of Tau Proteins
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On the day of immune staining, selected sections with LEnt on bregma −2.8 were washed in PBS for 10 min and then permeabilized in 0.5 ml of PBS/0.25% Triton X-100 (PBST). The tissues was blocked in 5% BSA and 5% normal donkey serum in PBST, 1.5 to 2 hours at RT. Then, we diluted primary antibodies in 5% BSA/PBST and incubated sections for overnight at 4°C. On the second day, brain sections were washed three times in PBST, 15 min each followed by incubation in secondary antibodies (1:700 for DyLight/Alexa-conjugated antibodies made in donkey purchased from Thermo Fisher Scientific) in 5% BSA/PBST and incubated for 2 hours at RT. For DAPI nucleus stain, DAPI was diluted (1:10,000) in PBST followed by incubation for 15 min. The brain sections were mounted onto microscope glass slides in ProLong Gold antifade reagent. The primary antibodies used in this study for immunofluorescence labeling are as follows: CP-13 (mouse; provided by P. Davies, Northwell), 1:300; TOMA2 (Anti-Tau oligomer clone 2, mouse,provided by Rakez Kayed laboratory), 1:200; FKBP12 (rat; R&D Systems, catalog no. MAB3777), 1:500.
3
Immunofluorescence Labeling of Hippocampus and LEnt
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For immuno-fluorescence labeling, selected sections of hippocampus from bregma −1.8 and LEnt from bregma −2.8, were washed in PBS for 10 mins and then permeabilized in 0.5ml PBS/0.25% Triton X-100 (PBST). Block tissues in blocking solution supplemented with 5% BSA and 5% normal donkey serum in PBST, 1.5–2 hours at RT. Dilute 1° antibodies in 5% BSA/PBST and incubate sections for overnight at 4°C. On the second day, wash the brain sections 3x in PBST, 15 min each. And then incubate the brain sections in 2° antibodies (1:700 for Dylight-/Alexa-conjugated antibodies made in donkey purchased from Thermo Fisher Scientific) in 5% BSA/PBST and incubate for 2 hours at RT. For DAPI nuclei stain, dilute DAPI (1:10,000) in PBST and incubate for 15 min. Wash 2x with PBST then 1x with PBS, 10 min each. Mount the brain sections onto microscope glass slides in Prolong gold anti-fade reagent. The primary antibodies used in this study are as follows: NeuN (chicken, Millipore, cat# ABN91), 1: 300; MC1 (mouse, provided by Peter Davies, Northwell), 1:100; TOMA2 (mouse, provided by Rakez Kayed, UTMB Galveston), 1:200 [26 (link)]; Images were captured by Carl Zeiss confocal LSM700.
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