Dissected brains were fixed with 4% paraformaldehyde for 15 min at room temperature. Fixed tissue was blocked with PBS containing 2% Triton X-100 and 10% normal goat serum (NGS) for 30 min and then incubated for 1 day each (with washing in between) at room temperature in PBS containing 1% Triton X-100, 0.25% NGS, and a primary antibody or secondary antibody solution. The brain was mounted in 20 μl Vectashield medium. We used the following primary antibodies at the indicated dilutions: 1:50 mouse anti-bruchpilot (nc82) (Developmental Studies Hybridoma Bank, Iowa), 1:1000 rabbit anti-5HT (Sigma, S5545). Secondary antibodies from Invitrogen were used at dilutions of 1:250, which were Alexa Fluor 633 goat anti-mouse lgG (Life Technologies, A21050), Alexa Fluor 568 goat anti-mouse lgG (Life Technologies, A11004), Alexa Fluor 633 goat anti-rabbit (Invitrogen, A21071). Confocal images were acquired with Zeiss LMS710 confocal laser scanning microscope under 40x or 63x magnification.
Alexa fluor 568 goat anti mouse lgg
Manufactured by Thermo Fisher Scientific
Alexa Fluor 568 goat anti-mouse IgG is a secondary antibody conjugated with the Alexa Fluor 568 fluorescent dye. It is designed to detect and visualize mouse immunoglobulin G (IgG) in various immunoassay techniques, such as Western blotting, immunohistochemistry, and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 633 goat anti rabbit, by Thermo Fisher Scientific (3 mentions)
Alexa fluor 633 goat anti mouse lgg, by Thermo Fisher Scientific (2 mentions)
Mouse anti bruchpilot nc82, by Developmental Studies Hybridoma Bank (2 mentions)
Alexa fluor 488 goat anti chicken, by Thermo Fisher Scientific (2 mentions)
Rabbit anti 5 ht, by Merck Group (1 mentions)
Lms710 confocal laser scanning microscope, by Zeiss (1 mentions)
Chicken anti gfp, by Thermo Fisher Scientific (1 mentions)
Rat anti n cadherin, by Developmental Studies Hybridoma Bank (1 mentions)
Fv1000, by Olympus (1 mentions)
Alexa fluor 647 donkey anti rat igg, by Abcam (1 mentions)
Lsm 710 microscope, by Zeiss (1 mentions)
Vectashield mounting medium, by Vector Laboratories (1 mentions)
3 protocols using alexa fluor 568 goat anti mouse lgg
1
Immunohistochemical Labeling of Drosophila Brains
2
Immunostaining Neuronal Markers in Drosophila
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We used the following primary antibodies at the indicated dilutions: 1:50 mouse anti-bruchpilot (nc82, Developmental Studies Hybridoma Bank, Iowa City, Iowa), 1:50 rat anti-CD8 Invitrogen (Waltham, MA)(MCD0800), 1:100 rabbit anti-5HT Sigma (S5545), 1:50 chicken anti-GFP Invitrogen (A10262), 1:200 rabbit anti-VAchT acbam (AB68984), and 1:200 Goat anti-ChAT EMD Millipore (AB144P). Secondary antibodies from Invitrogen were used at dilutions of 1:250, which were Alexa Fluor 633 goat anti-mouse lgG (Life Technologies, A21050), Alexa Fluor 568 goat anti-mouse lgG (Life Technologies, A11004), Alexa Fluor 633 goat anti-rabbit (Invitrogen, A21071), Alexa Fluor 488 goat anti-rat lgG (Life Technologies, A11006), Alexa Fluor 488 goat anti-chicken (Life Technologies, A11039), and Alexa Fluor 594 donkey anti-goat abcam (AB150136). Blocking serums were donkey serum from Sigma Aldrich (D9663) and goat serum Vector Labs (S-1000).
3
Synaptic Connectivity Mapping via syb:GRASP
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In brief, brains used for the induction of syb:GRASP were dissected and rinsed three times with a KCl solution (Macpherson et al., 2015 (link)). The brains were then fixed in 4% paraformaldehyde for 20 min. We used the following primary and secondary antibodies at the indicated dilutions: 1:1000 rabbit anti-5HT Sigma (S5545), 1:50 chicken anti-GFP Invitrogen (A10262), 1:100 mouse anti-GFP (referred as anti-GRASP, Sigma #G6539, ref:3), 1:500 rat anti-N-Cadherin (Developmental Studies Hybridoma Bank, DN-Ex #8), 1:250 Alexa Fluor 633 goat anti-rabbit (Invitrogen, A21071), 1:250 Alexa Fluor 488 goat anti-chicken (Life Technologies, A11039), 1:250 Alexa Fluor 568 goat anti-mouse lgG (Life Technologies, A11004) and 1:1000 Alexa Fluor 647 donkey anti-rat IgG (AbCam, ab150155). Brains were mounted and imaged in Vectashield mounting medium (Vector Labs). All steps were performed at room temperature. Confocal z-stacks for the syb:GRASP experiments were collected with a Zeiss LSM710 microscope using a 63 × oil immersion lens and the z-stack of GFP expression in Figure 1A was collected with an Olympus FV1000s using a 40x oil-immersion lens.
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