Reagents were purchased from Sigma-Aldrich (St. Louis, MO) unless otherwise indicated. The antibodies for cytokeratin-19 (CK-19) (ab 52625), desmin (ab185033) and apelin (ab 59469) were purchased from Abcam (Cambridge, MA); F4/80 (70076s), p-ERK (4370) and ERK (4695) were purchased from Cell Signaling Technology (Beverly, MA); α-SMA (α-smooth muscle actin, c6198) were purchased from Sigma-Aldrich; apelin receptor (sc-33823) was purchased from Santa Cruz (Dallas, TX). All selected primers were purchased from Qiagen (Germantown, MD); specific information about the primers was listed in Supplementary Table 1 . The iScript cDNA Synthesis Kit (170–8891) and iTaq Universal SYBR Green Supermix (172–5124) were purchased from Bio-Rad (Hercules, CA).
Ab59469
Manufactured by Abcam
Sourced in United States
Ab59469 is a primary antibody for use in immunoassays. It is a rabbit monoclonal antibody that targets the human Vimentin protein.
Automatically generated - may contain errors
Lab products found in correlation
Erk 4695, by Cell Signaling Technology (1 mentions)
Iscript cdna synthesis kit, by Bio-Rad (1 mentions)
Itaq universal sybr green supermix, by Bio-Rad (1 mentions)
Ab125213, by Abcam (1 mentions)
Ab69506, by Abcam (1 mentions)
Ab84296, by Abcam (1 mentions)
Ab6322, by Abcam (1 mentions)
Celltiter 96 aqueous one solution, by Promega (1 mentions)
Ab214369, by Abcam (1 mentions)
D9542, by Merck Group (1 mentions)
Tcs sp8, by Leica (1 mentions)
3 protocols using ab59469
1
Molecular Expression Analysis of Liver Cells
2
Apelin-13 Peptide Effects on Pericytes
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Exogenous recombinant apelin-13 peptide was purchased from Sigma (St. Louis, MO). The makers of pericytes were PDGFR-B (ab69506, Abcam, US), NG2 (SC-20162, Santa Cruz, CA), and Desmin (ab6322, Abcam, US). Antiapelin and anti-APJ were purchased from Abcam company (ab59469, ab125213, and ab84296, Abcam, US), respectively. CellTiter 96 AQueous One Solution was used in cell Proliferation Assay (Promega, US). Lentiviral vector knockout of apelin was constructed and purchased from GeneChem Co., Ltd. (Shanghai, China). Bcl-2 and Bax antibody were obtained from Cell Signaling Technology (#2870; #2772, CST, US).
3
Immunofluorescence Staining for Apelin and APJ
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All specimens were fixed with 4% paraformaldehyde (PFA), washed with PBS, permeabilized with 1% Triton-X 100 in PBS for 10 min, blocked with 1% BSA for 1 h at room temperature, and then incubated with primary antibodies overnight at 4℃. Specimens were then incubated with the fluorescent secondary antibodies and DAPI J o u r n a l P r e -p r o o f
(1:1000, D9542, Sigma) for 1 h in the dark at room temperature. The specimens were analyzed using a confocal microscope (LEICA TCS SP8). The primary antibodies used in immunofluorescence staining were as the follows: rabbit anti-Apelin (1:800, ab59469, Abcam), rabbit anti-APJ (1:800, ab214369, Abcam), mouse anti-myosin7α
(1:1000, DSHB), and rabbit anti-cleaved caspase-3 (1:1000, 9664S, Cell Signaling Technology).
(1:1000, D9542, Sigma) for 1 h in the dark at room temperature. The specimens were analyzed using a confocal microscope (LEICA TCS SP8). The primary antibodies used in immunofluorescence staining were as the follows: rabbit anti-Apelin (1:800, ab59469, Abcam), rabbit anti-APJ (1:800, ab214369, Abcam), mouse anti-myosin7α
(1:1000, DSHB), and rabbit anti-cleaved caspase-3 (1:1000, 9664S, Cell Signaling Technology).
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