Dulbecco's modified Eagle's medium (DMEM) and foetal bovine serum (FBS) were obtained from Gibco. Earle's balanced salt solution (EBSS) and propranolol were purchased from Sigma‐Aldrich. Rapamycin (RAPA), 3‐methyladenine (3‐MA) and chloroquine (CQ) were obtained from MedChemExpress, and ZVAD‐FMK, necrostatin‐1, liproxstatin‐1, SB203580, SP600125, SC79 and 740Y‐P were purchased from SelleckChem. Then, these reagents were dissolved in dimethyl sulfoxide (DMSO) (MP Biomedicals or water and stored at −80°C. Primary antibodies against Akt, p‐Akt, p38 MAPK, p‐p38 MAPK, JNK, p‐JNK, Erk1/2 and p‐Erk1/2 were purchased from Cell Signalling Technology. Primary antibodies against alpha‐smooth muscle actin (α‐SMA), fibronectin (FN), LC3B, P62, p‐PI3K p85, PI3K p85, ATG9b, ATG9a, mTOR, p‐mTOR, ATG12, ATG5 and anti‐ubiquitin were procured from Abcam. Primary antibodies against beta‐actin, alpha‐tubulin, GAPDH and HRP‐conjugated secondary antibodies were obtained from Proteintech. DyLight 549‐conjugated and DyLight 488‐conjugated secondary antibodies were provided by Abbkine.
Dylight 488 conjugated secondary antibodies
Manufactured by Abbkine
Sourced in United States
DyLight 488-conjugated secondary antibodies are fluorescent-labeled antibodies used for detecting and visualizing target proteins in various immunochemical applications, such as Western blotting, immunohistochemistry, and flow cytometry. These antibodies are conjugated with the DyLight 488 dye, which emits green fluorescence upon excitation, allowing for the specific detection of the target protein.
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Lab products found in correlation
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
P erk1 2, by Cell Signaling Technology (1 mentions)
Necrostatin 1, by Selleck Chemicals (1 mentions)
Hrp conjugated secondary antibody, by Proteintech (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Propranolol, by Merck Group (1 mentions)
Atg9a, by Abcam (1 mentions)
P pi3k p85, by Abcam (1 mentions)
P p38 mapk, by Cell Signaling Technology (1 mentions)
Dimethyl sulfoxide (dmso), by MP Biomedicals (1 mentions)
P mtor, by Abcam (1 mentions)
P jnk, by Cell Signaling Technology (1 mentions)
Anti ubiquitin, by Abcam (1 mentions)
Pi3k p85, by Abcam (1 mentions)
Z vad fmk, by Selleck Chemicals (1 mentions)
Earle s balanced salt solution ebss, by Merck Group (1 mentions)
Sp600125, by Selleck Chemicals (1 mentions)
Sb203580, by Selleck Chemicals (1 mentions)
Liproxstatin 1, by Selleck Chemicals (1 mentions)
Atg12, by Abcam (1 mentions)
2 protocols using dylight 488 conjugated secondary antibodies
1
Comprehensive Cell Signaling Reagents Protocol
2
Klotho Expression in T-Cell Lymphoma
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Primer sequence Klotho F, 5'-AGCAATCTGGTCTGAATAACACTGG-3' R, 5'-CATGTTTCAGCGTGAAAGTTCAAAG-3' GAPDH F, 5'-GGGAAACTGTGGCGTGAT-3' R, 5'-GAGTGGGTGTCGCTGTTGA-3' normal goat serum for 1 h. Then the slides were incubated with primary antibodies at 4˚C overnight. After washing with PBS, the DyLight 488-conjugated secondary antibodies (Abbkine, CA, USA) were added. Slides were washed and mounted with 4'6-diamino-2-phenylindole (DAPI; Boster, Wuhan, China).
Images were examined and recorded by Nikon IX73 fluorescent microscope.
Statistical analysis. All statistical analyses were performed by using statistic software SPSS17.0 (SPSS Inc., Chicago, IL, USA). Data are presented as the mean ± standard deviation (mean ± SD) from three separate experiments. Differences between groups were analyzed by one-way analysis of variance (ANOVA) or t-tests. P<0.05 was considered statistically significant. II. We next confirmed the expression of Klotho in T-cell lymphoma cell lines. As shown in Fig. 1B, T-cell lymphoma cells, exhibited remarkably lower mRNA levels of Klotho compared to the peripheral blood T lymphocytes from healthy donors. Decreased protein levels of Klotho expression were also noted in T-cell lymphoma cell lines (Fig. 1C).
Images were examined and recorded by Nikon IX73 fluorescent microscope.
Statistical analysis. All statistical analyses were performed by using statistic software SPSS17.0 (SPSS Inc., Chicago, IL, USA). Data are presented as the mean ± standard deviation (mean ± SD) from three separate experiments. Differences between groups were analyzed by one-way analysis of variance (ANOVA) or t-tests. P<0.05 was considered statistically significant. II. We next confirmed the expression of Klotho in T-cell lymphoma cell lines. As shown in Fig. 1B, T-cell lymphoma cells, exhibited remarkably lower mRNA levels of Klotho compared to the peripheral blood T lymphocytes from healthy donors. Decreased protein levels of Klotho expression were also noted in T-cell lymphoma cell lines (Fig. 1C).
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