For analysis of the indirect immunofluorescence assay, a mixture of MERS-CoV-infected and non-infected Vero cells at a ratio of 3:1 was seeded onto slide glasses. The cells were then fixed with acetone and incubated with the normal mouse IgG, 492-1G10E4E2 or 506-2G10G5 monoclonal antibody at 37°C for 2 h. The samples were further incubated with Alexa Flour 488-conjugated goat anti-mouse IgG antibody (Thermo Fisher Scientific Co.). Finally, the samples were mounted and analyzed using a fluorescence microscope (1×70, Olympus Co.) (28 (link), 29 (link)). To visualize the confocal microcopy, Vero cells (5 × 104) were seeded onto cover glasses in 12 well plates and infected with MERS-CoV (0.1 MOI). After two days, the infected cells were fixed with 4% paraformaldehyde and subsequently blocked with 1% BSA and 0.1% triton X-100 in PBS. The slides were incubated in the presence of the 492-1G10E4E2 or 506-2G10G5 monoclonal antibody for 2 h, washed and then incubated with the Alexa Flour 488-conjugated goat anti-mouse IgG antibody for 1 h. Hoechst 33258 (Thermo Fisher Scientific) was used to stain the nuclei. The slides were examined by Carl Zeiss LSM710 (Carl Zeiss Co. Ltd.).
Alexa flour 488 conjugated goat anti mouse igg antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Alexa Fluor 488-conjugated goat anti-mouse IgG antibody is a secondary antibody that binds to mouse immunoglobulin G (IgG) antibodies. It is conjugated with the Alexa Fluor 488 fluorescent dye, which emits green fluorescence when excited by an appropriate light source. This antibody is commonly used in immunofluorescence techniques to detect and visualize target proteins in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710, by Zeiss (2 mentions)
Hoechst 33258, by Thermo Fisher Scientific (2 mentions)
Carl lsm710, by Zeiss (2 mentions)
Fv200 microscope, by Olympus (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Alexa flour 488 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
4 protocols using alexa flour 488 conjugated goat anti mouse igg antibody
1
Indirect Immunofluorescence Assay for MERS-CoV Detection
2
Imaging of Plasmodium vivax Schizonts
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The schizont-stage-rich parasites of P. vivax were collected from malaria patients in Thailand, as described previously [18 (link)]. Briefly, the slides were blocked with PBS containing 5% nonfat milk, incubated with rabbit anti-MSP1-19 (1:200 dilution) [18 (link)] and mouse anti-MSP8 (1:100 dilution) as primary antibodies, followed by incubation with Alexa Flour 546-conjugated goat anti-rabbit IgG or Alexa Flour 488-conjugated goat anti-mouse IgG antibody (Invitrogen, Carlsbad, CA, USA) and nuclear staining with DAPI (Invitrogen). Then, the slides with ProLong Gold antifade reagent (Invitrogen) were mounted. The parasites were observed under oil immersion using a confocal laser scanning FV200 microscope (Olympus, Tokyo, Japan).
3
Intracellular Localization of MUC1-C
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Cells were cultured on poly-L-lysine-coated glass cover slips in 12-well culture plates. After cells were cultured for 48 h, cells were fixed with 4% paraformaldehyde for 10 min. For detection of cell surface MUC1-C, mouse anti-hMUC1 monoclonal antibody, anti-MUC1-CT2 antibody, or mouse normal IgG were treated for 2 h on ice. For intracellular staining, cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with 3% BSA, and stained with anti-hMUC1 monoclonal antibody for 2 h at room temperature. After washing in PBS-T containing 1% BSA, the samples were incubated with Alexa Flour-conjugated secondary antibody such as Alexa Flour 488-conjugated goat anti-mouse IgG antibody (for anti-hMUC1 monoclonal antibody, Invitrogen) or Alexa Flour 594-conjugated goat anti-Armenian hamster IgG antibody (for anti-MUC1-CT2 antibody, Invitrogen) for 1 h. Nuclei were stained with Hoechst 33258. All samples were mounted and observed using the confocal laser scanning microscope system (CLSM system; LSM 710, Carl Zeiss, Jena, Germany) 30 (link).
4
MERS-CoV Indirect Immunofluorescence Assay
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To perform the indirect immunofluorescence assay (IFA), Vero cells were mixed with MERS-CoV infected Vero cells (ratio 1:3) and plated onto slide glasses. The mixed cells were fixed with acetone, washed with distilled water (DW), and incubated with normal mouse IgG or the MERS-M158 peptide-specific monoclonal antibody at 37 °C. After a 2 h incubation, the slides were washed with PBS and DW, and then incubated with Alexa Flour 488-conjugated goat anti-mouse IgG antibody (Thermo Fisher Scientific, Waltham, MA, USA). The samples were mounted and then observed using a fluorescence microscope (IX70, Olympus, Tokyo, Japan). To obtain confocal images, Vero cells (5 × 104 cells) were plated onto cover glass in 12-well culture plates and infected with MERS-CoV (0.1 MOI) for 2 days. The cells were treated with 4% paraformaldehyde to fix the cells and incubated with PBS containing 1% BSA and 0.1% triton X-100 for 30 min. to block the non-specific binding. The MERS-M158 peptide-specific monoclonal antibody was loaded into the wells and incubated for 2 h and then incubated with Alexa Flour 488-conjugated goat anti-mouse IgG for 1 h. The nuclei were observed by Hoechst 33258 (Thermo Fisher Scientific) staining. The cells were examined by Carl Zeiss LSM710 (Carl Zeiss, Oberkochen, DE).
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