After lineage differentiation, cells were washed twice with PBS and fixed with 4 % formalin for 10 min at room temperature. For Oil red O staining, fixed cells were then stained with Oil red O reagent (Beyotime) for 1 h at room temperature; for Alizarin red staining, fixed cells were stained with 1% Alizarin red (Leagene, Beijing, China) with pH 4.2 for 30 min at room temperature; for Alcian Blue staining, fixed cells were then colored for 30 min in a 1% Alcian Blue solution (OriCell). Finally, cells were washed with water for three times to remove unbound dye and photographed under light microscopy.
Alizarin red
Manufactured by Leagene
Sourced in China
Alizarin red is a dye commonly used as a histological stain for the detection of calcium deposits in tissues. It binds to calcium ions, creating a bright red color that can be observed under a microscope. This product is used in various research and analytical applications that require the identification of calcium-containing structures.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Oil red o, by Beyotime (1 mentions)
Paraformaldehyde (pfa), by Leagene (1 mentions)
Hematoxylin, by Abcam (1 mentions)
Paraformaldehyde (pfa), by Solarbio (1 mentions)
Hoechst dye, by Thermo Fisher Scientific (1 mentions)
Bcip nbt alkaline phosphatase color development kit, by Leagene (1 mentions)
Sirius red, by Leagene (1 mentions)
Alamar blue, by Thermo Fisher Scientific (1 mentions)
Sodium hyaluronate, by Lifecore Biomedical (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
β glycerophosphate, by Merck Group (1 mentions)
10 protocols using alizarin red
1
Multilineage Differentiation Staining
2
Alizarin Red Staining of Calcification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells in culture were washed twice with PBS, fixed in 4% paraformaldehyde for 10 min, and then washed with distilled water and stained with 1% Alizarin red (Leagene, Beijing, China) for 30 min at room temperature. Then, cells were washed with distilled water to remove the unbound dye, and calcification was visualized as red deposits under light microscopy.
3
Alizarin Red Staining for Calcium Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
alizarin red staining was performed to detect calcium salt deposition on days 12 or 15 after the initiation of osteogenic differentiation. In brief, cells were washed twice with PBS, fixed with 4% paraformaldehyde for 10 min, rinsed with double-distilled H2O, and stained with 1% alizarin red (pH 4.2; Leagene, Beijing, China) solution for 30 min at room temperature. Cells were then washed with double-distilled water to remove the unbound dye and were imaged by light microscopy.
4
Bone Mineralization Visualization Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ALP staining was performed using the ALP Staining Kit (Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences, Tianjin, China). Alizarin red staining was performed to detect matrix mineralization deposition during the later stage of osteogenesis as described27 (link). In brief, cells in culture were washed twice with phosphate-buffered saline (PBS), fixed in 4% paraformaldehyde for 10 min, and then washed with distilled water and stained with 1% Alizarin red (Leagene, Beijing, China) for 30 min at room temperature. Then, cells were washed with distilled water to remove the unbound dye, and matrix calcification was visualized as red deposits under light microscopy.
5
Alizarin Red Staining of Osteoblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Osteoblasts were inoculated into 24-well plates and cultured with DMEM containing 10% FBS to a density of 5 × 105 cells per well. The medium was discarded, and the osteoblasts were washed twice with phosphate-buffered saline (PBS). The osteoblasts were fixed with 10% formaldehyde-calcium solution for 10 min and washed with isopropanol for 1 min. The osteoblasts were stained with alizarin red (Leagene, Beijing, China) for 1 min at 37 °C in a dark room. After decolorization, the osteoblasts were counterstained with hematoxylin (Abcam, Cambridge, MA, USA) for 1 min. Finally, the slices were washed with PBS and mounted with glycerin. Then, the slices were visualized under a microscope.
6
Alkaline Phosphatase and Alizarin Red Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Alkaline phosphatase (ALP) staining was performed using an ALP staining kit (Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences, Tianjin, China) according to the manufacturer’s protocol. Alizarin red staining was performed to detect matrix mineralization deposition during the later stage of osteogenesis. Briefly, cells were washed twice with PBS, fixed in 4% paraformaldehyde for 10 min, rinsed with distilled water and stained with 1% Alizarin red (Leagene, Beijing, China) with pH 4.2 for 30 min at room temperature. Then, cells were rinsed with distilled water to remove the unbound dye and matrix calcification was shown with red deposition.
7
Cytokine-Induced Mineralization Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Alizarin Red staining (ARS) was carried out to determine the effects of cytokines on mineral depositions. After 7, 14, and 28 days of induction, the cultures were fixed in 4% paraformaldehyde (Solarbio) for 30 minutes, stained with Alizarin Red (Leagene) for 30 minutes, and then desorbed with cetylpyridinium chloride (Aladdin, China) with continuous agitation. The absorbance at 560 nm of representative cultures was measured.
8
Alizarin Red Staining of Mineralized Bone Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Alizarin Red staining was applied to measure matrix mineralization deposition in MC3T3-E1 cells [14 ]. In brief, DEX-induced osteoporosis cells were fixed in 4% paraformaldehyde (aladdin, Shanghai, China) for 10 min, and then washed with distilled water and stained with 1% Alizarin Red in accordance with the manufacturer’s protocol (Leagene, Beijing, China). Finally, the stained cells were rinsed with distilled water, matrix calcification was observed, and red deposition were observed under the microscope (Nikon, Tokyo, Japan).
9
Osteoblast Differentiation Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ALP staining was performed using an ALP staining kit (Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences, Tianjin, China) according to the manufacturer’s protocol on days 4 and 6 of osteoblast differentiation. alizarin red staining was conducted to detect matrix mineralization deposition on days 12 and 15 after the initiation of differentiation. In brief, cells were washed twice with PBS, fixed with 4% paraformaldehyde for 10 min, rinsed with double-distilled H2O, and stained with 1% alizarin red (pH 4.2; Leagene, Beijing, China) staining solution for 30 min at room temperature. The cells were photographed following a thorough wash in double-distilled H2O to remove the unbound dye.
10
Titanium-Coated Silicon Wafer for Bone Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Titanium-coated silicon wafer (0.5×0.5 cm) was produced by vapor deposition as a gift from the Interdisciplinary Nanoscience Center (iNANO). CS (150 kDa, 95% deacetylation) was provided by HEPPE MEDICAL (Halle, Germany). Sodium hyaluronate (Mw ≈360 kDa) was purchased from Life-core Biomedical, LLC (Chaska, MN, USA). Polyethylenimine ([PEI] Mw 750 kDa, 50 wt% solution), β-glycerophosphate, ascorbic acid, and dexamethasone were obtained from Sigma-Aldrich Co. (St Louis, MO, USA). siRNA duplex targeting murine TNF-α (siTNF) sequence (sense, 5′-pGU-CUCAGCCUCUUCUCAUUCCUGct-3′, antisense, 5′-AG-CAGGAAUGAGAAGAGGCUGAGACAU-3′) and siGFP sequence (sense, 5′-GACGUAAACGGCCACAAGUUC-3′, antisense: 5′-ACUUGUGGCCGUUUACGUCGC-3′) were bought from Dharmacon (Lafayette, CO, USA). siRNA duplex targeting human Ckip-1 sequence (sense, 5′-CGCACAGUCAGUACCGGAAdT*dT-3′, antisense, 5′-UUCCGGUACUGACUGUGCGdT*dT-3′) was ordered from GenePharma (Shanghai, People’s Republic of China). Hoechst dye, RiboGreen® kit, Lipofectamine® 2000, and alamarBlue® were bought from Thermo Fisher Scientific (Waltham, MA, USA). BCIP/NBT Alkaline Phosphatase Color Development Kit, Sirius Red, and Alizarin Red were bought from Leagene (Beijing, People’s Republic of China).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!