The following combinations of antibodies were used for doublelabeling immunohistochemistry: anti-Col VI and anti-Runx2; anti-NG2 and anti-Runx2; and anti-PCNA and anti-Runx2. The antigen-antibody reactivities were detected with a mix of Alexa Fluor 488-conjugated anti-rabbit IgG chicken antibody (1 : 400; Invitrogen, Carlsbad, CA, USA) and Alexa Fluor 594-conjugated anti-goat IgG donkey antibody (1 : 400; Invitrogen), or Alexa Fluor 488-conjugated anti-mouse IgG donkey antibody (1 : 400; Invitrogen) and Alexa Fluor 594-conjugated anti-goat IgG donkey antibody (1 : 400; Invitrogen). Double-labeling immunohistochemistry with anti-NG2 and anti-Col VI antibodies (both raised in rabbit) was performed according to the protocol outlined by Jackson ImmunoResearch Laboratories. After the sections were incubated with anti-NG2, the immunoreactivity was detected with an excess amount of Alexa Fluor 488-conjugate anti-rabbit IgG donkey Fab fragment (H+L; 1 : 60; Jackson ImmunoResearch, West Grove, PA, USA). After washing in PBS, the sections were incubated with anti-Col VI antibody, followed by treatment with Alexa Fluor 594-conjugated anti-rabbit IgG goat antibody (1 : 400; Invitrogen). The sections were examined with DP2-BSW software (Olympus, Tokyo, Japan).
Alexa fluor 594 conjugated donkey anti goat igg antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 594-conjugated donkey anti-goat IgG antibody is a secondary antibody used to detect and visualize primary goat antibodies in various immunoassay applications. It is conjugated with the Alexa Fluor 594 fluorescent dye, which can be detected using appropriate fluorescence detection methods.
Automatically generated - may contain errors
Lab products found in correlation
Dp2 bsw software, by Olympus (1 mentions)
Pcdna3, by Thermo Fisher Scientific (1 mentions)
Effectene, by Qiagen (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 conjugated donkey anti rabbit igg antibody, by Thermo Fisher Scientific (1 mentions)
Vectashield, by Vector Laboratories (1 mentions)
Fv1000, by Olympus (1 mentions)
3 protocols using alexa fluor 594 conjugated donkey anti goat igg antibody
1
Immunohistochemistry of Extracellular Matrix and Progenitor Cells
2
MERS-CoV Susceptibility Evaluation in Transfected Cells
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MERS-CoV-resistant PESU-B5L, R05T, R06E, or Tb1Lu or MERS-CoV-susceptible EidNi/41.3, EpoNi/22.1, HypLu/45.1, HypNi/1.1, RoNi/7.1, RoNi/7.2, or Vero E6 cells were transfected with a plasmid expressing human CD26/DPP4 (pCMV-xL-hDPP4, Origene Technologies, Rockville, MD) or control plasmid pcDNA3.1+ (Life Technologies) by Effectene (Qiagen, Frederick, MD) or Lipofectamine 3000 (Life Technologies) according to the manufacturer's instruction. At 24 h or 48 h post transfection, cells were washed once with 0% DMEM and then inoculated with MERS-CoV/EMC at an MOI of 3. Bat cells were incubated at 37°C for 1 h with gently rocking of the plates every 15 min. At 1 h after exposure, cells were washed twice with 0% DMEM, and 0.5 ml of 2% DMEM was added. At 24 h post-exposure, supernatants were harvested for virus yield determination. Plates were fixed with 10% NBF. Plates were stained with goat anti-human CD26/DPP4 followed by Alexa Fluor 594-conjugated donkey anti-goat IgG antibody and/or polyclonal rabbit anti-MERS-CoV spike protein antibody followed by Alex Fluor 488-conjugated chicken anti-rabbit IgG antibody (Life Technologies). Images were acquired using the Operetta high content imaging system.
3
Immunofluorescence Localization of HRP and Collagen IV
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Some deparaffinized tissue sections were blocked with 5% fish gelatin in PBS for 1 hr and treated with rabbit anti-HRP and goat anti-collagen type IV polyclonal antibodies at 4°C overnight (Fig. 1 n). Then, they were incubated with Alexa Fluor 488-conjugated donkey anti-rabbit IgG antibody and Alexa Fluor 594-conjugated donkey anti-goat IgG antibody (Life Technologies, Eugene, OR, USA) at room temperature for 1 hr. Finally, intracellular nuclei were counterstained with 4,6-diamidino-2-phenylidole (DAPI), and the immunostained sections were mounted on glass slides with Vectashield (Vector) and observed under a confocal laser scanning microscope (FV-1000; Olympus) (Fig. 1 p).
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