Immunofluorescence experiments were performed as previously described [22 (link)]. Lung tissue slices were incubated with anti-BiP (11587-1-AP, Proteintech) and anti-vimentin antibodies (BF8006, Affinity Biosciences, Changzhou, China). Slices containing climbing fibers were incubated with anti-vimentin (BF8006, Affinity Biosciences) and anti-E-cadherin (3195 T, CST, MA, USA) antibodies. After incubation with secondary antibodies and DAPI, the slices were observed under a fluorescence microscope (Olympus, Japan).
Bf8006
Manufactured by Affinity Biosciences
Sourced in United States
The BF8006 is a laboratory equipment product manufactured by Affinity Biosciences. It is a precision scientific instrument designed for specialized applications in research and analysis. The core function of the BF8006 is to perform specific tasks within a controlled laboratory environment, but a detailed description of its intended use cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Af1032, by Affinity Biosciences (1 mentions)
Ripa lysate solution, by Beyotime (1 mentions)
Af6191, by Affinity Biosciences (1 mentions)
Af6032, by Affinity Biosciences (1 mentions)
Af4009, by Affinity Biosciences (1 mentions)
Af6308, by Affinity Biosciences (1 mentions)
Af6423, by Affinity Biosciences (1 mentions)
Af3423, by Affinity Biosciences (1 mentions)
Af3308, by Affinity Biosciences (1 mentions)
Wgpvdf22, by Wuhan Servicebio Technology (1 mentions)
Af5128, by Affinity Biosciences (1 mentions)
Af5384, by Affinity Biosciences (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Cy3 conjugated goat anti rabbit igg, by Wuhan Servicebio Technology (1 mentions)
G1202, by Wuhan Servicebio Technology (1 mentions)
Bf9212, by Affinity Biosciences (1 mentions)
Fitc conjugated goat anti mouse igg, by Wuhan Servicebio Technology (1 mentions)
G1012, by Wuhan Servicebio Technology (1 mentions)
Fv3000, by Olympus (1 mentions)
3 protocols using bf8006
1
Immunofluorescence Imaging of Lung Tissue
2
Western Blot Analysis of Cell Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Preprocessed samples were added to the RIPA lysate solution (P0013B; Beyotime, China), and the supernatants were collected after centrifugation. Samples were loaded onto sodium dodecyl sulfate-polyacrylamide gels (3250GR500; neoFROXX, Einhausen, Germany). Proteins were transferred to polyvinylidene difluoride membranes (WGPVDF22; Servicebio, China) after electrophoresis in transfer buffer for 5 min. The membranes were then incubated with primary antibodies (1:5000 dilution) targeting CD31 (AF6191, Affinity, USA), Vimentin (BF8006, Affinity), α-SMA (AF1032, Affinity), slug (350,136, ZENBIO, China), snail (AF6032, Affinity), twist (AF4009, Affinity), LC3II (3868S, CST, USA), p62 (AF5384, Affinity), beclin 1 (AF5128, Affinity), AMPK (AF6423, Affinity), p-AMPK (AF3423, Affinity), mTOR (AF6308, Affinity), and p-mTOR (AF3308, Affinity). Horseradish peroxidase-conjugated secondary antibodies at a 1:10,000 dilution were added after washing. The membranes were then viewed with an automatic darkroom exposure instrument (JS-M6P; P&Q, China) and varying luminous intensities were used for optimal exposure.
3
Immunohistochemical Analysis of Cellular Markers
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The paraffin‐embedded tissue slides were dewaxing and antigen retrieval utilizing citric acid buffer (10 mM, pH 6.0; Servicebio, G1202). Following this, block the glass slides with BSA and incubate overnight at 4°C with the specified primary antibody. The primary antibody employed was as follow: anti‐CCL20 antibody (1:200; Affinity, DF2238); anti‐CD4 antibody (1:500; Elabscience, E‐AB‐22098); anti‐CD116 antibody (1:500; Affinity, DF4820); anti‐alpha SMA (α‐SMA) antibody (1:500; Affinity, BF9212); anti‐vimatin antibody (1:500; Affinity, BF8006). Subsequently, the slides were left at room temperature for 2 h with the appropriate secondary antibodies, which were as follows: Cy3‐conjugated Goat Anti‐Rabbit IgG (1:500; Servicebio, GB21303); FITC‐conjugated Goat Anti‐Mouse IgG (1:200; Servicebio, GB22301). The marked slides were always protected from light and were dyed using DAPI (Servicebio; G1012). The co‐localization of CCL20 with other biomarkers was recorded using confocal laser scanning microscopy (Olympus; FV3000). The fluorescence signal was quantified using ImageJ.
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