Bs 0295g cy3

Manufactured by Bioss Antibodies

Sourced in China

Bs-0295G-Cy3 is a fluorescently labeled secondary antibody that can be used in various immunoassay applications. The Cy3 fluorescent dye is conjugated to the secondary antibody, providing a way to detect and visualize target proteins or molecules in a sample.

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Lab products found in correlation

5 protocols using bs 0295g cy3

Immunostaining of Goat Spermatogonial Stem Cells

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In this study, we used promyelocytic leukemia zinc finger (PLZF) and Thy-1 cell surface antigen (THY1) as specific markers of goat SSCs for immunofluorescence staining. Briefly, the cells were pipetted into a 96-well plate (3599; Corning) precoated with poly-L-lysine (P2100; Solarbio), with approximately 5 × 10³ cells per well. Then, the cells were fixed in 4% paraformaldehyde (PFA; P1110; Solarbio) for 15 min and then permeabilized by the addition of 0.25% Triton X-100 (T8200; Solarbio). Thereafter, 10% of goat serum was added for blocking. A diluted primary antibody (1 : 200; bs-5971R and bs-0778R; Bioss, Beijing, China) was added and incubated overnight at 4°C. After rinsing with PBST, a diluted fluorescent secondary antibody (1 : 200; bs-0295G-Cy3 and bs-0295G-FITC; Bioss) was added and incubated for 1 h at room temperature. After incubation, the cells were rinsed three times with PBST. We added 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI; C0060; Solarbio), and the cells were rinsed after 5 min of incubation. The cells were observed under a fluorescence microscope (Nikon, Tokyo, Japan).

Feng T.Y., Li Q., Ren F., Xi H.M., Lv D.L., Li Y, & Hu J.H. (2020). Melatonin Protects Goat Spermatogonial Stem Cells against Oxidative Damage during Cryopreservation by Improving Antioxidant Capacity and Inhibiting Mitochondrial Apoptosis Pathway. Oxidative Medicine and Cellular Longevity, 2020, 5954635.

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Immunofluorescence Analysis of AKT and ST7L

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Cells cultured on glass coverslips were fixed with 4% paraformaldehyde, blocked with 5% bovine serum albumin and incubated with AKT (pan) antibody (no.4691; Cell Signaling Technology) and ST7L antibody (sc-138649; Santa Cruz, Biotechnology) overnight at 4 °C. Then, cells were incubated with mouse anti-goat IgG/FITC antibody (bs-0294M-FITC, BIOSS) and goat anti-rabbit IgG/Cy3 antibody (bs-0295G-Cy3, BIOSS) for 2 h at room temperature. DAPI (Invitrogen) was used to stain the nucleus. LEICA DMI4000B microscope (Leica, Heidelberg, Germany) was used to take the images.

Zhuang L., Wang X., Wang Z., Ma X., Han B., Zou H., Wu Z., Dong S., Qu Z., Zang Y, & Wu L. (2017). MicroRNA-23b functions as an oncogene and activates AKT/GSK3β/β-catenin signaling by targeting ST7L in hepatocellular carcinoma. Cell Death & Disease, 8(5), e2804-.

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Immunostaining of Synaptic Proteins

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Formalin-fixed and paraffin-embedded tissue chip samples were obtained from Tianjin Genink Biotechnology Co. Ltd. (Tianjin, China). After dewaxing, antigen repairing, and spontaneous fluorescence quenching, paraffin sections were permeabilized with Triton X-100 for 30 min, blocked with 1% bovine serum albumin (BSA) for 30 min, and incubated with primary antibodies overnight at 4 °C. The sections were washed with phosphate-buffered saline (PBS) 3 times for 5min each time and then the secondary antibodies were added and incubated for 50 min in the dark at room temperature. Nuclei were stained with DAPI and incubated for 10 min in the dark at room temperature. The sections were sealed with antifade solution and stored at 4 °C in the dark, and images were photographed using a fluorescence microscope (BX51, Olympus, Tokyo, Japan). The primary antibodies used were synaptophysin (1:100; ab32127, Abcam, Cambridge, UK) and Homer1 (1:100, ab211415, Abcam). The secondary antibodies used were goat anti-rabbit IgG-CY3 (1:200, BS-0295G-CY3; Bioss, Beijing, China) and goat anti-rabbit IgG-FITC (1:200, BS-0295G-FITC; Bioss).

Li K., Liang X., Xie X., Tian L., Yan J., Lin B., Liu H., Lai W., Liu X, & Xi Z. (2023). Role of SHANK3 in concentrated ambient PM2. 5 exposure induced autism-like phenotype. Heliyon, 9(3), e14328.

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Immunofluorescence Microscopy of HepG2 Cells

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Before cell seeding, a small 8 mm × 8 mm slide was placed in every well in a 24-well plate. When HepG2 cells approach approximately 60% confluence, cell slides were washed in PBS and fixed in 95% ethylalcohol for 30 min at room temperature. The cell slides were then incubated with primary antibody for 12 h at 4 ℃, followed by a brief wash thrice with PBS for 5 min and 1 h incubation at 37 ℃ with secondary fluorescent antibodies, IgG/Cy3 (bs-0295G-Cy3, Bioss, Beijing) and IgM/RBITC (bs-0368R-RBITC, Bioss, Beijing). Slides were rinsed in PBS for 5 min three times, and nucleus was stained with 1 mg/mL DAPI of 200 µL (Beyotime Biotechnology) for 15 min. After drying, the mounted slides were pictured with a fluorescence microscope. The mean immunofluorescent intensity was calculated for target proteins using the software ImageJ.

Li Q., Wang S.J., Wang W.J., Ye Y.C., Ling Y.Q, & Dai Y.F. (2023). PAK4-relevant proliferation reduced by cell autophagy via p53/mTOR/p-AKT signaling. Translational Cancer Research, 12(3), 461-472.

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Immunofluorescence Analysis of Isolated CAFs and NFs

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The isolated CAFs or NFs were fixed in 4% paraformaldehyde. After permeabilization in 0.2% Triton X-100 and blocking in goat serum, the cells were probed with primary antibodies α-SMA (1:500, bsm-33187M, Bioss, Beijing, China) and vimentin (1:100, bsm-0756R, Bioss) overnight at 4°C. Subsequently, Goat Anti-Mouse IgG/FITC (1:100, bs-0296G-FITC, Bioss) and Goat Anti-rabbit IgG/Cy3 (1:100, bs-0295G-Cy3, Bioss) were applied. After nuclear counterstaining with DAPI, the cells were observed using a fluorescent microscope (Olympus, Tokyo, Japan).

Fan J.T., Zhou Z.Y., Luo Y.L., Luo Q., Chen S.B., Zhao J.C, & Chen Q.R. (2021). Exosomal lncRNA NEAT1 from cancer-associated fibroblasts facilitates endometrial cancer progression via miR-26a/b-5p-mediated STAT3/YKL-40 signaling pathway. Neoplasia (New York, N.Y.), 23(7), 692-703.

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