Alkaline phosphatase alp staining kit
Manufactured by Solarbio
Sourced in China
The Alkaline phosphatase (ALP) staining kit is a laboratory reagent used to detect and visualize the presence of the ALP enzyme in biological samples. ALP is an enzyme involved in various physiological processes and its expression can indicate specific cellular activities. The kit provides the necessary components to perform ALP staining, allowing for the identification and localization of ALP-positive cells or tissues.
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Lab products found in correlation
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Cdna synthesis kit, by Takara Bio (2 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (2 mentions)
D gal, by Merck Group (2 mentions)
Bcl 2, by Cell Signaling Technology (2 mentions)
Malonaldehyde mda and superoxide dismutase sod assay kit, by Nanjing Jiancheng (2 mentions)
Antibody to β actin, by Proteintech (2 mentions)
Second antibody conjugated hrp goat anti mouse and goat anti rabbit, by Proteintech (2 mentions)
Goldner trichrome staining kit, by Solarbio (2 mentions)
Tartrate resistant acid phosphatase staining kit, by Solarbio (2 mentions)
3 methyladenine 3 ma, by Selleck Chemicals (2 mentions)
Runx2, by Cell Signaling Technology (2 mentions)
Oil red o staining kit, by Solarbio (2 mentions)
Dm2500, by Leica (1 mentions)
Alizarin red kit, by Solarbio (1 mentions)
Alizarin red, by Solarbio (1 mentions)
5 protocols using alkaline phosphatase alp staining kit
1
Osteogenic Differentiation Assay Protocol
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d -Gal and GlcN were procured from Sigma-Aldrich (Darmstadt, Germany). 3-Methyladenine (3-MA) was purchased from Selleckchem (Houston, TX, USA). ELISA kits for cytokines IL-1β and IL-6 were procured from Boster (Wuhan, Hubei, China). Malonaldehyde (MDA) and superoxide dismutase (SOD) assay kit was purchased from Jiancheng Bioengineering (Nanjing, Jiangsu, China).The cDNA was synthesized by reverse transcription using a cDNA synthesis kit (Takara, Japan), and was analyzed with SYBR Green Real-time PCR kit (Novizan Biotechnology, Nanjing, Jiangsu, China).The antibodies included LC3, Beclin-1, P62, RUNX2, OCN, BMP2, Bcl-2, BAX and Cleaved Caspase-3 were purchased from Cell Signaling Technology (Danvers, MA, USA), and the antibodies to Ki-67 and p16 were all obtained from Beyotime (Shanghai, China). The antibody to β-actin and the second antibody conjugated HRP (goat-anti-mouse and goat-anti-rabbit) were purchased from Proteintech (Chicago, IL, USA). The Alkaline phosphatase (ALP) staining kit, Goldner trichrome staining kit and Tartrate-resistant acid phosphatase (TRAP) staining kit were purchased from Solarbio (Beijing, China).
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2
5-HT Induced Osteogenic Differentiation
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BMSCs were seeded at a density of 2 × 105 cells/well in 6‐well plates. After the cell confluence reached 80%, the cells were treated with 10−5 M 5‐HT for 3 days. The experiment was then performed osteogenic induction for 14 days. Alkaline phosphatase (ALP) staining kit (Solarbio) was used to qualitatively detect the expression of ALP. Alizarin red staining (ARS) staining kit (Solarbio) was used to visualize the mineralization module formation of 5‐HT intervention. The cells were observed under a light microscope (Olympus) to evaluate the degree of osteogenic capability.
3
Adipocyte and Osteoblast Staining Protocol
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The cells obtained after dissolution [32 (link)] of the matrices were resuspended in PBS. The cell suspension was evenly spread on the adherent glass slide, which was then dried at 60°C for 8 min. An oil red O staining kit (Solarbio, China) was used to stain adipocytes, while an alkaline phosphatase (ALP) staining kit (Solarbio, China) was used to stain osteoblasts. An optical microscope (Leica, DM2500, Germany) was used to obtain images.
4
Comprehensive Cell Staining Protocols
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ALP staining: After 10–14 days of osteogenic induction, the BMSCs were fixed in 4% formaldehyde for 5 min at room temperature. In accordance with the protocol, alkaline phosphatase (ALP) staining kit (G1480, Solarbio, Beijing, China) was used for incubating with working solutions for 20 min. After the staining solution is removed by washing with PBS twice, the cells are dried in the air. An inverted microscope (DMI1, Leica, Wetzlar, Germany) was used to observe the staining results.
Alizarin red staining: After 21 days of osteogenic induction, the cells were fixed with 95% ethanol for 10 min and stained according to the instructions of Alizarin red kit (G1450, Solarbio, Beijing, China). The stained calcium nodules were observed under an optical microscope.
Oil red O staining: After seven days of adipogenic induction, the cells were fixed with 10% neutral formalin for 1 h and stained according to the instructions of Oil red O staining kit (G1262, Solarbio, Beijing, China). After washing with distilled water, the stained lipid droplets were observed under an inverted microscope.
Alizarin red staining: After 21 days of osteogenic induction, the cells were fixed with 95% ethanol for 10 min and stained according to the instructions of Alizarin red kit (G1450, Solarbio, Beijing, China). The stained calcium nodules were observed under an optical microscope.
Oil red O staining: After seven days of adipogenic induction, the cells were fixed with 10% neutral formalin for 1 h and stained according to the instructions of Oil red O staining kit (G1262, Solarbio, Beijing, China). After washing with distilled water, the stained lipid droplets were observed under an inverted microscope.
5
Osteoporosis Intervention via Autophagy
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D-Gal and GlcN were procured from Sigma-Aldrich (Darmstadt, Germany). 3-Methyladenine (3-MA) was purchased from Selleckchem (Houston, TX, USA). ELISA kits for cytokines IL-1β and IL-6 were procured from Boster (Wuhan, Hubei, China). Malonaldehyde (MDA) and superoxide dismutase (SOD) assay kit was purchased from Jiancheng Bioengineering (Nanjing, Jiangsu, China).The cDNA was synthesized by reverse transcription using a cDNA synthesis kit (Takara, Japan), and was analyzed with SYBR Green Real-time PCR kit (Novizan Biotechnology, Nanjing, Jiangsu, China).The antibodies included LC3, Beclin-1, P62, RUNX2, OCN, BMP2, Bcl-2, BAX and Cleaved Caspase-3 were purchased from Cell Signaling Technology (Danvers, MA, USA), and the antibodies to Ki-67 and p16 were all obtained from Beyotime (Shanghai, China). The antibody to β-actin and the second antibody conjugated HRP (goat-anti-mouse and goat-anti-rabbit) were purchased from Proteintech (Chicago, IL, USA). The Alkaline phosphatase (ALP) staining kit, Goldner trichrome staining kit and Tartrate-resistant acid phosphatase (TRAP) staining kit were purchased from Solarbio (Beijing, China).
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