Immunofluorescent staining of ISH-stained tissues and paraffin sections was performed as described previously (12 (link), 23 (link)). Primary antibodies were diluted as follows: 1:200 rabbit anti-CDH1 (3195, Cell Signaling Technology, Beverly, MA), 1:250 mouse anti-CDH1 (610181, BD Transduction Laboratories, San Jose, CA) 1:50 mouse anti-KRT14 (ms-115-p0, Thermo Fisher Scientific), 1:250 rabbit anti-AR (sc-816, Santa Cruz Biotechnology, Santa Cruz, CA), 1:200 rabbit anti-DNMT1 (5032, Cell Signaling Technology). Secondary antibodies were diluted as follows: 1:250 Dylight 549-conjugated goat anti-rabbit IgG (111-507-003, Jackson ImmunoResearch, West Grove, PA), 1:250 Dylight 488-conjugated goat anti-rabbit IgG (111-487-003, Jackson ImmunoResearch) and 1:250 Dylight 488-conjugated goat anti-mouse IgG (115-487-003, Jackson ImmunoResearch). Immunofluorescently labeled tissues were counterstained with 4’,6-diamidino-2-phenylindole, dilactate (DAPI), and mounted in antifade medium (phosphate buffered saline containing 80% glycerol and 0.2% n-propyl gallate). Whole mount immunohistochemistry was performed as described previously (23 (link)). Primary antibody was diluted 1:750 rabbit anti-CDH1 and secondary antibody was diluted 1:500 biotin conjugated goat anti-rabbit IgG (BA-1000, Vector, Burlingame, CA).
Dylight 488 conjugated goat anti rabbit igg
Manufactured by Jackson ImmunoResearch
Sourced in Panama, United States
DyLight 488-conjugated goat anti-rabbit IgG is a secondary antibody used for detection and visualization in immunoassays. It is produced by immunizing goats with rabbit IgG and conjugating the resulting antibodies with the DyLight 488 fluorescent dye.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710 confocal microscope, by Zeiss (3 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Rabbit anti cdh1, by Cell Signaling Technology (1 mentions)
Mouse anti cdh1, by BD (1 mentions)
Dylight 549 conjugated goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Biotin conjugated goat anti rabbit igg ba 1000, by Vector Laboratories (1 mentions)
Dylight 488 conjugated goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)
M20008, by Abmart (1 mentions)
A11120, by Thermo Fisher Scientific (1 mentions)
Ta180004, by OriGene (1 mentions)
Dylight 488 conjugated affinipure goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Dylight 594 conjugated affinipure goat anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Rabbit anti s100, by Merck Group (1 mentions)
Cy3 conjugated goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Mouse anti beta 3 tubulin, by Promega (1 mentions)
P s6ser240 244, by Cell Signaling Technology (1 mentions)
Volocity, by Quorum Technologies (1 mentions)
4 protocols using dylight 488 conjugated goat anti rabbit igg
1
Immunofluorescent Staining of ISH-Stained Tissues
2
Multicolor Immunostaining of Embryos
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Embryos were fixed in 4% paraformaldehyde overnight, then rinsed with PBST four times every 5 min. Next, the resulting embryos were blocked at room temperature for at least 1 h in PBST containing 10% heat-inactivated goat serum and 1% BSA, and then stained using the following affinity-purified primary antibodies overnight at 4°C: anti-ZsYellow (1:200; 632475, Clontech), anti-ZsYellow (1:800; TA180004, Origene), anti-Cdh5 (1:200; 555289, BD Pharmingen), anti-pERK1/2 (1:1000; 9101, Cell Signaling), anti-pAKT (1:400; 4060, Cell Signaling Technology), anti-Scl (1:200; NBP2-50285, Novus Biologicals), anti-Etv2 (1:500; ES1004, Kerafast), anti-GFP (1:1000; A11120, Invitrogen) and anti-Flag (1:500; M20008, Abmart). Samples were then washed for 3-4 h with PBST, followed by incubation with secondary antibodies for 1 h at room temperature, including DyLight 488-conjugated goat anti-rabbit IgG (1:200; 711-545-152, Jackson), DyLight 594-conjugated goat anti-mouse IgG (1:200; 715-585-150, Jackson), DyLight 488-conjugated AffiniPure goat anti-mouse IgG (1:200; 715-545-150, Jackson) and DyLight 594-conjugated AffiniPure goat anti-rabbit IgG (1:200; 711-585-152, Jackson). DAPI (1:10,000, Sigma) was used as a nuclear stain.
3
Multimodal Immunocytochemical Staining Protocol
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For whole-gel differential immunostaining, cultures were fixated for 2 h in 4% paraformaldehyde (PFA) solution in PBS, and then permeabilized and blocked with 0.3–1% Triton and 4% fetal bovine serum (FBS) solution in PBS (4 h). The cultures were then washed with PBS, and incubated (overnight, 4°C) with the following primary antibodies: Mouse-Anti beta III tubulin (1:400, Promega), a neuronal cell marker, and Rabbit-Anti S100 (1:200, Sigma), a marker of glial cells. Cultures were then washed with PBS and exposed (4 h to overnight) to CY3-conjugated goat-anti-mouse IgG (1:100, Jackson), Dylight 488-conjugated goat-anti-rabbit IgG (1:100, Jackson), and DAPI (6 nM, Sigma), designed to stain the nuclei of all cells and incubated for at least 4 h, before being washed again with PBS. Cells were imaged using a confocal microscope (Zeiss, LSM 700).
4
Phosphorylated S6 Protein Imaging in Mouse Brain
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