The treated BEAS-2B cells were fixed with 4% paraformaldehyde and permeated with 0.1% Triton X-100 for 10 min. After incubation with anti-HMGB1 at 4 °C overnight, the cells were stained with secondary Alexa Fluor®488-conjugated antibody (Beyotime Biotechnology, Shanghai, China) for 1 h at 37 °C in the dark. The nuclei were counterstained with DAPI. Images were visualized by using a fluorescence microscope.
Alexa fluor 488 conjugated antibody
Manufactured by Beyotime
Sourced in China
The Alexa Fluor®488-conjugated antibody is a fluorescently labeled antibody that can be used for various applications, such as immunofluorescence, flow cytometry, and fluorescence microscopy. The Alexa Fluor®488 dye is a bright, green-fluorescent dye that can be excited by a 488 nm laser and emits light at around 520 nm. The conjugation of this dye to the antibody allows for the specific detection and visualization of target proteins or cells.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 555 conjugated antibody, by Thermo Fisher Scientific (1 mentions)
Optiphot 2 microscope, by Nikon (1 mentions)
α actinin, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Nis element, by Nikon (1 mentions)
Paraformaldehyde (pfa), by Beyotime (1 mentions)
5 protocols using alexa fluor 488 conjugated antibody
1
HMGB1 Immunofluorescence Staining
2
Immunofluorescent Labeling of S100a9
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Cells were immersed in medium containing 3.7% paraformaldehyde for 1 h, then permeabilized with 0.5% Triton X-100, and blocked with normal goat serum. Anti-S100a9 antibody (26992-1-AP; Proteintech) was added and incubated for 12 h at 4°C. Then, Alexa Fluor 488-conjugated antibody (Beyotime) was added and incubated for 1 h. Images were observed using a fluorescence microscope.
3
Immunofluorescent Staining of Cardiac Cells
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Cells were first fixed in 4% paraformaldehyde (Beyotime Institute of Biotechnology) for 15 min at room temperature (RT), permeabilized with 0.1% Triton X-100 (Sigma-Aldrich: Merck KGaA) for 10 min at RT and blocked with 10% goat serum (Sigma-Aldrich; Merck KGaA) at RT for 1 h. Samples were subsequently stained with primary antibodies against cardiac troponin T (cTnT; 1:200; cat. no. 710635; Invitrogen; Thermo Fisher Scientific, Inc.) and α-actinin (Invitrogen; 1:200; cat. no. 710947; Thermo Fisher Scientific, Inc.), dissolved in PBS at 4°C for 12 h and further incubated with Alexa Fluor™ 555-conjugated antibody (1:500; cat. no. A0460; Beyotime Institute of Biotechnology) and Alexa Fluor™ 488-conjugated antibody (1:500; cat. no. A0423; Beyotime Institute of Biotechnology) at RT for 2 h. Nuclei were stained using 1 µg/ml DAPI (Sigma-Aldrich; Merck KGaA) at RT for 5 min. An Optiphot-2 microscope (Nikon Corporation) equipped with a CCD video camera system (Optronics Engineering, Ltd.) and a computer interface (NIS-Elements; version 4.2.0; Nikon Corporation) was used for imaging analysis (magnification, ×100). In total, 40 images were taken for each well.
4
Immunofluorescence and Cell Viability Assay
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The treated BEAS-2B cells were xed with 4% paraformaldehyde and permeated with 0.1% Triton X-100 for 10 min. After incubation with anti-HMGB1 at 4℃ overnight, the cells were stained with secondary Alexa Fluor®488-conjugated antibody (Beyotime Biotechnology, Shanghai, China) for 1 hr at 37℃ in the dark. The nuclei were counterstained with DAPI. Images were visualized by using a uorescence microscope.
Cell Viability Assay BEAS-2B cells were seeded into a 96-well plate at a density of 3000 cells/well and treated as described above. Then cell viability was assessed with the Cell Counting Kit-8 assay (CCK8; Dojindo Laboratories, Tokyo, Japan) at 0,12,24,48hr, respectively. The optical density(OD) values of the absorbance at 450nm were measured using Riorad microplate reader.
Cell Viability Assay BEAS-2B cells were seeded into a 96-well plate at a density of 3000 cells/well and treated as described above. Then cell viability was assessed with the Cell Counting Kit-8 assay (CCK8; Dojindo Laboratories, Tokyo, Japan) at 0,12,24,48hr, respectively. The optical density(OD) values of the absorbance at 450nm were measured using Riorad microplate reader.
5
Immunofluorescence and Cell Viability Assay
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The treated BEAS-2B cells were xed with 4% paraformaldehyde and permeated with 0.1% Triton X-100 for 10 min. After incubation with anti-HMGB1 at 4℃ overnight, the cells were stained with secondary Alexa Fluor®488-conjugated antibody (Beyotime Biotechnology, Shanghai, China) for 1 hr at 37℃ in the dark. The nuclei were counterstained with DAPI. Images were visualized by using a uorescence microscope.
Cell Viability Assay BEAS-2B cells were seeded into a 96-well plate at a density of 3000 cells/well and treated as described above. Then cell viability was assessed with the Cell Counting Kit-8 assay (CCK8; Dojindo Laboratories, Tokyo, Japan) at 0,12,24,48hr, respectively. The optical density(OD) values of the absorbance at 450nm were measured using Riorad microplate reader.
Cell Viability Assay BEAS-2B cells were seeded into a 96-well plate at a density of 3000 cells/well and treated as described above. Then cell viability was assessed with the Cell Counting Kit-8 assay (CCK8; Dojindo Laboratories, Tokyo, Japan) at 0,12,24,48hr, respectively. The optical density(OD) values of the absorbance at 450nm were measured using Riorad microplate reader.
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