After deparaffinization and rehydration of FFPE liver sections according to a standard protocol, antigen retrieval was conducted by boiling in 10 mM sodium citrate buffer. After cooling down, the slides were permaebilized and blocked using 0.4% Triton X-100 (BioBasic) and 5% BSA (Capricorn) in PBS, consecutively. After that, the slides were incubated with an anti-LC3B antibody (Thermo Fisher Scientific) overnight at 4 °C. After twice washing with PBS (10 minutes per wash), the slides were further incubated with a Dylight 488 conjugated anti-rabbit secondary antibody (Thermo Fisher Scientific) for 2 hours in the dark at room temperature. Finally, the slides were washed and mounted by using Permount (Merck) for further analysis under fluorescence microscope.
Anti lc3b antibody
Manufactured by Thermo Fisher Scientific
The Anti-LC3B antibody is a laboratory reagent used for the detection and quantification of the LC3B protein, a key marker of autophagy, in various biological samples. This antibody specifically binds to the LC3B protein, allowing researchers to analyze its expression and dynamics during cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Capricorn (1 mentions)
Permount, by Merck Group (1 mentions)
Triton x 100, by Bio Basic (1 mentions)
Phenazopyridine, by Merck Group (1 mentions)
Gefitinib, by Merck Group (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Pik93, by Merck Group (1 mentions)
Anti phospho akt, by Cell Signaling Technology (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions)
Fluorescence microscope, by Nikon (1 mentions)
3 protocols using anti lc3b antibody
1
FFPE Liver Immunostaining for LC3B
2
Cellular Signaling Pathway Analysis
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Phenazopyridine, PIK93, NIH12848, and gefitinib were purchased from Sigma. UNC3230 was obtained from RnDSystems. The anti-LC3B antibody was bought from Thermofisher, and anti-AKT and anti-phospho AKT were purchased from Cell Signaling.
3
Immunofluorescence Assay for LC3B
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C-33A cells were fixed in 4% formaldehyde at room temperature for 10 min, before being washed twice with PBS buffer solution. 0.5% Triton X-100 was adopted to permeabilize cells. After being blocked with 5% bovine serum albumin (Thermo Fisher Scientific, Inc.) for 1 h at room temperature, cells were incubated with anti-LC3B antibody (1:50; cat. no. ab192890; Abcam) at 4˚C overnight. After washing with PBS, cells were incubated with Alexa Fluor 488-conjugated secondary antibody (1:200; cat. no. ab150077; Abcam) at room temperature for 2 h. The nuclei were stained by DAPI (0.01 mg/ml; Vector Laboratories, Inc.). The images were observed under a fluorescence microscope (Nikon Corporation).
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