The alkaline phosphatase staining was performed using a BCIP/NBT Alkaline Phosphatase Color Development Kit (Leagene, Beijing, China). For immunofluorescence, the cells that were cultured in human PSCeasy Medium (Cellapy, Beijing, China) were harvested and fixed with phosphate-buffered saline (PBS) and paraformaldehyde (4%) for 15 min at room temperature. For the molecules localized in the nucleus, the cells were treated with Triton X-100 (0.5%) for 15 min and with BSA (3%) for 30 min. Then, the cells were incubated overnight at 4 °C in BSA (3%) with the primary antibodies and were washed with PBS 3 times. Then, the cells were incubated for 60 min at 37 °C in BSA (3%) with the secondary antibodies against the pluripotency markers (Cellapy, Beijing, China). The nuclei were counterstained by DAPI, and the images were taken by an Olympus fluorescence microscope (BX51) (Olympus, Tokyo, Japan).
Bcip nbt alkaline phosphatase color development kit
Manufactured by Leagene
Sourced in China
The BCIP/NBT Alkaline Phosphatase Color Development Kit is a laboratory reagent used for the detection and visualization of alkaline phosphatase activity in various biological applications, such as immunohistochemistry, Western blotting, and enzyme-linked immunosorbent assays (ELISA).
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Human psceasy medium, by Cellapy (1 mentions)
Stereomicroscope, by Olympus (1 mentions)
Ix73 inverted fluorescence microscope, by Olympus (1 mentions)
Hoechst dye, by Thermo Fisher Scientific (1 mentions)
Alizarin red, 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)
3 protocols using bcip nbt alkaline phosphatase color development kit
1
Pluripotency Marker Immunofluorescence Staining
2
Macrophage-Conditioned Media Effects on Osteoblast Differentiation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collection and preparation of conditioned medium (CM). RAW 264.7 cells were cultured in a growth medium supplemented with HA, Au-HA, and PDA@Au-HA for 3 days. The culture supernatants were collected and diluted with α-MEM containing 10% FBS at a ratio of 1:1 before use.
Alkaline Phosphatase (ALP) Analysis. MC3T3-E1 cells were seeded in a cell culture dish with 1.5 × 10 5 cells per dish. The macrophage-CM (HA, Au-HA, and PDA@Au-HA) was refreshed every 3 days. ALP staining was performed using a BCIP/NBT alkaline phosphatase color development kit (LEAGENE, China) on the 7th day, and photographic images were captured via a stereomicroscope (Olympus, Japan).
Alizarin Red S (ARS) Staining. The MC3T3-E1 cells were cultured with different macrophage-CM for 14 days in the culture dish. The cells were preserved for 20 min at room temperature using 4% paraformaldehyde. After three PBS washes, the cells were stained for ten minutes with 1% (w/v) Alizarin Red S (pH = 4.2) (Haixing, China). We used an Olympus IX73 inverted fluorescence microscope to view the samples stained with ARS.
Alkaline Phosphatase (ALP) Analysis. MC3T3-E1 cells were seeded in a cell culture dish with 1.5 × 10 5 cells per dish. The macrophage-CM (HA, Au-HA, and PDA@Au-HA) was refreshed every 3 days. ALP staining was performed using a BCIP/NBT alkaline phosphatase color development kit (LEAGENE, China) on the 7th day, and photographic images were captured via a stereomicroscope (Olympus, Japan).
Alizarin Red S (ARS) Staining. The MC3T3-E1 cells were cultured with different macrophage-CM for 14 days in the culture dish. The cells were preserved for 20 min at room temperature using 4% paraformaldehyde. After three PBS washes, the cells were stained for ten minutes with 1% (w/v) Alizarin Red S (pH = 4.2) (Haixing, China). We used an Olympus IX73 inverted fluorescence microscope to view the samples stained with ARS.
3
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!