Rabbit polyclonal anti-Aβ42 antibody was procured from Millipore (Millipore, Cat# AB5078P, CA, USA). Rabbit polyclonal anti-amylin antibody was obtained from Boster (Boster, Cat#A00414, Wuhan, China). Mouse monoclonal anti-Glu4 (Santa Cruz, Cat# sc-53566, CA, USA) and Na+/K+-ATPases antibodies were purchased from Santa Cruz (Santa Cruz, Cat# sc-58628, CA, USA). Rabbit polyclonal anti-GAPDH antibody was purchased from ImmunoWay (ImmunoWay, Cat# YM3445, TX, USA). β-Galactosidase staining kits (Beyotime, Cat# C0602, Haimen, China) and membrane and cytosol protein extraction kits were purchased from Beyotime (Beyotime, Cat# P0027, Haimen, China).
β galactosidase staining kit
Manufactured by Beyotime
Sourced in China
The β-galactosidase staining kit is a laboratory tool used to detect the presence and activity of the β-galactosidase enzyme. The kit provides the necessary reagents and protocol for performing this staining procedure, which is commonly used in cellular and molecular biology applications.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin, by Thermo Fisher Scientific (2 mentions)
Lsm 710, by Zeiss (2 mentions)
Anti gapdh antibody, by Immunoway (1 mentions)
Primary antibodies against lc3b, by Cell Signaling Technology (1 mentions)
Type 2 collagen, by Cell Signaling Technology (1 mentions)
Beclin 1, by Cell Signaling Technology (1 mentions)
Immunofluorescence reagents, by Beyotime (1 mentions)
Collagenase type 2, by Merck Group (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Western blotting reagents, by Beyotime (1 mentions)
Inverted fluorescence microscope, by Olympus (1 mentions)
β galactosidase, by Beyotime (1 mentions)
Inverted bright field microscope, by Leica (1 mentions)
Masson trichrome staining, by Solarbio (1 mentions)
Pannoramic midi, by 3DHISTECH (1 mentions)
Bx53 microscope, by Olympus (1 mentions)
Ab259341, by Abcam (1 mentions)
Ab133626, by Abcam (1 mentions)
Ab124962, by Abcam (1 mentions)
48 protocols using β galactosidase staining kit
1
Antibody Procurement and Protein Extraction
2
Chondrocyte Autophagy and Differentiation
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Alcian dye, rat-derived recombinant PTH and type II collagenase were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). Primary antibodies against LC3b, Beclin-1, P62, ATG5, type II collagen, SOX-9 and β-actin were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). β-galactosidase staining kits, immunofluorescence reagents, western blotting reagents, DAPI, and cell protein extraction kits were purchased from Beyotime Institute of Biotechnology (Shanghai, China). Finally, cell culture reagents, including Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS) and 0.25% trypsin were purchased from Gibco (Thermo Fisher Scientific, Inc., Waltham, MA, USA).
3
Senescence Assay Protocol
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The cells were plated in 6-well plates at approximately 30% confluence. After pretreatment, the medium was removed, and the cells were washed once with PBS. Then, 1 mL of β-galactosidase staining fixative was added, and the cells were fixed at room temperature for 15 min. The cells were then washed three times with PBS, the mixture was added according to the instructions of the β-galactosidase staining kit (Beyotime, Shanghai, China), and the cells were incubated in the absence of carbon dioxide overnight at 37°C. Finally, images were obtained under an inverted fluorescence microscope (Olympus, Tokyo, Japan).
4
Senescence-associated β-galactosidase Assay
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The senescent cells were identified with a β‐galactosidase Staining Kit (Beyotime), which uses X‐gal as the substrate and produces dark blue products under the catalysis of aging‐specific β‐galactosidase. Blue cells expressing β‐galactosidase can easily be observed under a light microscope. The cells were stained with β‐galactosidase, fixed at room temperature for 10 min and washed with 0.01 M PBS (Beyotime) three times for 5 min each. Then, a working dye solution comprising a mixture of β‐galactosidase staining solution A (10 μl), β‐galactosidase staining solution B (10 μl), β‐galactosidase staining solution C (930 μl), and X‐gal (50 μl) was added, and the mixture was incubated at 37°C for 4 h. The samples were observed, photographed and counted under an ordinary optical microscope. If photos were not taken sufficiently quickly, the dye solution was discarded, an equivalent volume of PBS was added, and the cells were stored at 4°C for several days. The proportion of positive cells was determined as the number of β‐galactosidase positive cells in 10 fields divided by the total number of cells and was compared through statistical analysis between groups.
5
Senescence Induction in Endothelial Progenitor Cells
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Long-term culture was used to establish the senescent model of EPCs. In brief, EPCs were cultured in 6-well plate not coated (vehicle group) or coated with biomaterial membranes (HA∗-Col I, HA-Col I∗, CS∗-Col I, or CS-Col I∗) for 4 days. After that, β-Galactosidase Staining Kit (Beyotime, China) was used to analyze the senescent EPCs. In brief, after washing with PBS, EPCs grown on 6-well plate not coated (vehicle group) or coated with biomaterial membranes (HA∗-Col I, HA-Col I∗, CS∗-Col I, or CS-Col I∗) were treated with 2% formaldehyde and 0.2% glutaraldehyde in PBS for 6 min and then incubated with fresh X-gal staining solution (1 mg/mL X-gal, 5 mmol/L potassium ferrocyanide, 5 mmol/L potassium ferricyanide, and 2 mmol/L MgCl2; pH 6) for 12 h at 37°C without CO2. After staining, blue-stained cells and total cells were counted at 3 different microscopic fields. The percentage of β-galactosidase positive cells was calculated.
6
Measuring β-galactosidase Activity in Cells
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β-galactosidase activity was examined by β-galactosidase staining kit according to manufacturer's instructions (Beyotime, China). Briefly, cells were fixed and rinsed with PBS. Then fixed cells were added β-galactosidase liquid substrate for 24-48 h. Finally, cells were rinsed in deionized water for several times.
7
Senescence Evaluation via β-Galactosidase Staining
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β-galactosidase staining kit (Beyotime, Shanghai, China) was employed to indicate the senescence. The protocol was executed as recommended by the kit instruction.
8
Cellular Senescence Measurement via β-Galactosidase Staining
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Cellular senescence was assessed using a β-Galactosidase Staining Kit (Beyotime). Briefly, culture media was removed and cells were washed with PBS, cells were fixed with 4% paraformaldehyde (PFA, v/v) at RT for 15 min. Fixation solution was removed and cells were rinsed 3 times with PBS for 5 min each. Cells were then treated with 1 mL staining solution and incubated overnight at 37 °C. β-Galactosidase staining was then observed using a bright field inverted microscope (Leica).
9
Renal β-galactosidase Activity Evaluation
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The expression of β-galactosidase (SA-β-gal) in renal tissues was analyzed with a β-galactosidase staining kit (Beyotime, Shanghai, China) in accordance with the manufacturer’s protocols.
10
Senescence β-Galactosidase Activity Assay
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Senescence β-Galactosidase (SA- β-Gal) activity was evaluated with a β-galactosidase staining kit from Beyotime (#C0602). For the tissue staining, equilibrate the frozen tissue to room temperature. Rinse the tissue with PBS 3 times for 5 min each time. Add an appropriate amount of β-galactosidase Staining Fixative to fully cover the tissue and incubate at room temperature for 15 min. And then rinse the tissue with PBS 3 times for 5 min each time. Remove the PBS, add an appropriate amount of staining working solution. Incubate overnight at 37℃, and the nest day, examine by a light microscope.
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