Paraformaldehyde phosphate buffer

Manufactured by Fujifilm

Sourced in Japan

Paraformaldehyde phosphate buffer is a laboratory reagent used to prepare fixative solutions for microscopy and histology applications. It is a commonly used fixative that preserves the structure and composition of biological samples. The buffer helps maintain the pH and ionic environment necessary for effective fixation.

Automatically generated - may contain errors

Lab products found in correlation

Osteogenesis assay kit, by Merck Group (3 mentions) 1 step nbt bcip plus suppressor solution, by Thermo Fisher Scientific (3 mentions) Mesenchymal cell growth supplement, by Lonza (1 mentions) β glycerophosphate, by Lonza (1 mentions) Penicillin streptomycin, by Lonza (1 mentions) L glutamine, by Lonza (1 mentions) Dexamethasone (dex), by Lonza (1 mentions) Pe goat anti mouse igg, by BioLegend (1 mentions) Facscanto 2 flow cytometer, by BD (1 mentions) Aniline blue orange g solution, by Fujifilm (1 mentions) Phosphotungstic acid, by Fujifilm (1 mentions) Cm3050, by Leica (1 mentions) α mem, by Fujifilm (1 mentions) Biorevo bz 9000, by Keyence (1 mentions) Slowfade gold antifade reagent, by Thermo Fisher Scientific (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Coverslip, by Matsunami (1 mentions)

9 protocols using paraformaldehyde phosphate buffer

Alkaline Phosphatase and Alizarin Red Staining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Alkaline phosphatase histochemistry was performed at 1 d, 3 d, 5 d, 7 d, 10 d, 2 w, and 3 w during the induction culture [31]
[36]
[37] . When the ALP staining was done, the medium was removed, and the cell layers were rinsed with PBS 3 times and fixed in 4% Paraformaldehyde Phosphate Buffer (Wako Pure Chemical Industries, Ltd.) for 5 minutes at room temperature. After 5 minutes at room temperature, the cell layers were washed with deionized water. Then, the fixed cells were incubated with 1-Step NBT/BCIP plus Suppressor Solution (Thermo Fisher Scientific INC.). After 30 minutes incubation at 37°C, the cell layers were washed with deionized water and observed both grossly and with the light microscope.
Before Alizarin Red staining, the cell layers were rinsed with PBS 3 times and fixed in 4% Paraformaldehyde Phosphate Buffer (Wako Pure Chemical Industries, Ltd.) for 10 minutes at room temperature. Then the cells were washed with deionized water for 3 times. Then the Alizarin red staining was done following the standard protocol as described in the Osteogenesis Assay Kit (ECM815, Millipore) [38] .

Fang X., Murakami H., Demura S., Hayashi K., Matsubara H., Kato S., Yoshioka K., Inoue K., Ota T., Shinmura K, & Tsuchiya H. (2014). A Novel Method to Apply Osteogenic Potential of Adipose Derived Stem Cells in Orthopaedic Surgery. PLoS ONE, 9(2), e88874.

+ Open protocol

+ Expand

Alkaline Phosphatase and Alizarin Red Staining

Check if the same lab product or an alternative is used in the 5 most similar protocols

Alkaline phosphatase histochemistry was performed at 1 d, 3 d, 5 d, 7 d, 10 d, 2 w, and 3 w during the induction culture period. When the ALP staining was done, the medium was removed, and the cell layers were rinsed with PBS 3 times and fixed in 4% Paraformaldehyde Phosphate Buffer (Wako Pure Chemical Industries, Ltd.) for 5 minutes at room temperature. After 5 minutes at room temperature, the cell layers were washed with deionized water. Then, the fixed cells were incubated with 1-Step NBT/BCIP plus Suppressor Solution (Thermo Fisher Scientific INC.). After 30 minutes incubation at 37°C, the cell layers were washed with deionized water and observed both grossly and with the light microscope.
Before being Alizarin Red stained, the ADSCs cell layers were rinsed with PBS 3 times and fixed in 4% Paraformaldehyde Phosphate Buffer (Wako Pure Chemical Industries, Ltd.) for 10 minutes at room temperature. Then the cells were washed with deionized water for 3 times. Then the Alizarin red staining was done following the standard protocol as described in the Osteogenesis Assay Kit (ECM815, Millipore).

+ Open protocol

+ Expand

Osteogenic Differentiation of hPDLMSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Induction of osteogenic differentiation and the alizarin red staining procedure were performed in accordance with a previously published protocol [17 (link)]. For osteogenic differentiation, 4.0 × 10³ monolayer-cultured hPDLMSCs, two hPDLMSC spheroids (2000 cells/spheroid), or two co-cultured spheroids (hPDLMSCs: HUVECs = 1:1, 1:2, and 2:1) (2000 cells/spheroid) were seeded in a 48-well cell culture plate (Iwaki) as monolayer conditions in culture medium, and the culture medium was changed to hMSC Osteogenic Induction Medium (OIM) containing dexamethasone, L-glutamine, ascorbate, penicillin/streptomycin, mesenchymal cell growth supplement, and β-glycerophosphate (Lonza) when cells reached confluent. At 7, 10, 13, 17, and 21 days after medium change, the cells were fixed with a 4% paraformaldehyde phosphate buffer (Wako). Deposited calcium was stained with a 1% alizarin red solution (Wako) and quantified using ImageJ (National Institutes of Health, Bethesda, MD, USA). The alizarin red positive colony ratio was calculated from the bottom area of the wells and the area of nodules stained with alizarin red.

Sano K., Usui M., Moritani Y., Nakazawa K., Hanatani T., Kondo H., Nakatomi M., Onizuka S., Iwata T., Sato T., Togari A., Ariyoshi W., Nishihara T, & Nakashima K. (2020). Co-cultured spheroids of human periodontal ligament mesenchymal stem cells and vascular endothelial cells enhance periodontal tissue regeneration. Regenerative Therapy, 14, 59-71.

+ Open protocol

+ Expand

Quantitative Flow Cytometry of Cell Surface Antigen Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

When the MT-2 and Jurkat cell cultures reached a density of 1 × 10⁶ cells/mL, 10 μg/mL mAbs (clone A–D) were added for 16 h at 4 °C. Next, the cells were washed twice with wash buffer [containing 2% (v/v) FBS and 0.02% (w/v) NaN₃ in PBS] and incubated with phycoerythrin (PE) goat anti-mouse IgG (BioLegend, San Diego, CA, USA) as a secondary antibody for 1 h on ice under light. Next, the cells were washed twice with wash buffer. Finally, the cells were fixed in 4% (v/v) paraformaldehyde phosphate buffer (FUJIFILM Wako Pure Chemical Corporation) and subjected to fluorescence-activated cell sorting (FACS) analysis using a BD FACS Canto TM II Flow Cytometer (BD Biosciences, Franklin Lakes, NJ, USA).

Hatayama Y., Yamaoka Y., Morita T., Jeremiah S.S., Miyakawa K., Nishi M., Kimura Y., Mitsunaga M., Iwase T., Kimura H., Yamamoto N., Takaori-Kondo A., Hasegawa H, & Ryo A. (2022). Development of a Monoclonal Antibody Targeting HTLV-1 Envelope gp46 Glycoprotein and Its Application to Near-Infrared Photoimmuno-Antimicrobial Strategy. Viruses, 14(10), 2153.

+ Open protocol

+ Expand

Histological Analysis of Bone and Cementum Formation

Check if the same lab product or an alternative is used in the 5 most similar protocols

The maxillae were dissected and fixed with a 4% paraformaldehyde phosphate buffer (Wako) for 3 days. The maxilla blocks were decalcified in Morse's solution at 4 °C for 4 days, followed by dehydration and paraffin embedding. Serial tissue sections with thickness of 8 μm were cut out and stained with haematoxylin-eosin (H&E). The area of newly formed bone and the rate of new cementum length were measured using ImageJ, as described previously [31 (link),32 (link)]. The area of newly formed bone was estimated as the ratio of the area surrounded by the top of the newly formed bone and reference notches on the mesiobuccal and palatal root surfaces. The newly formed cementum was defined as the portion of cementum that was found on the crown side above the notches, and in which Sharpey's fibers were inserted. The rate of new cementum length was defined as the ratio of the new cementum length formed on the root surface facing the defect to the root surface length from notch to notch. Regarding the cell transplant groups, Azan staining was used to confirm the running of Sharpey's fibers. Paraffin-embedded samples were stained with 0.1% Azocarmin G (Wako), Aniline alcohol (Wako), Acetic acid-ethanol (Wako), 5% Phosphotungstic acid (Wako), and Aniline Blue-Orange G Solution (Wako).

+ Open protocol

+ Expand

Kidney Tissue Harvesting and Preservation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mice were anesthetized, and after confirming the absence of righting reflex, the abdomen was incised. After perfusion with phosphate-buffered saline (PBS) using a pump, the kidneys were harvested and placed in 4% paraformaldehyde/phosphate buffer (pH 7.0, FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) at 4 °C overnight.

Tsuji T., Hosoda A., Toriyama Y., Yoshida Y, & Kohno T. (2022). Renin-angiotensin system inhibitors combined with cisplatin exacerbate cisplatin-induced nephrotoxicity in mice. Translational Oncology, 18, 101369.

+ Open protocol

+ Expand

Tissue Fixation and Cryosectioning of Mouse Maxillary Bone

Check if the same lab product or an alternative is used in the 5 most similar protocols

A catheter was inserted into the heart of 8-week-old mice that had been euthanized with CO₂ gas. After purging with PBS, the mice were reflux-fixed in 4% paraformaldehyde phosphate buffer (Wako Pure Chemical Corporation), and the maxillary bone was removed. The maxillary bone was subsequently immersed overnight in PBS containing 20% sucrose. We prepared the frozen sections at a thickness of 20 µm from OCT blocks (Sakura Seiki, Tokyo, Japan) after they had been placed in a freezer at − 80 °C for 10 min; they were then in Leica CM3050 S (Leica, Nussloch, Germany) at − 20 °C for 10 min.

Miki K., Takeshita N., Yamashita M., Kitamura M, & Murakami S. (2024). Calcitonin gene-related peptide regulates periodontal tissue regeneration. Scientific Reports, 14, 1344.

+ Open protocol

+ Expand

Osteogenic Differentiation of Antibiotic-loaded ADSCs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibiotic-loaded ADSCs and ADSCs were analysed for their capacity for osteogenic differentiation using ALP staining and alizarin red histochemistry. To induce differentiation, cells were cultured in osteogenic medium composed of α-MEM (Wako Pure Chemical Industries) containing 10% FBS, 0.1 mM dexamethasone, 50 mM ascorbate-2-phosphate, 10 mM β-glycerophosphate, and 1% penicillin-streptomycin^{52 (link)}. Alkaline phosphatase (ALP) histochemistry was performed at 2 weeks after osteogenic induction culture. For ALP staining, cells were rinsed with PBS three times and fixed in 4% paraformaldehyde phosphate buffer (Wako) at room temperature for 5 min. They were then washed with deionized water. The fixed cells were incubated with 1-Step NBT/BCIP plus Suppressor Solution (Thermo Fisher Scientific) at 37 °C for 30 min, washed with deionized water, and observed both with the naked eye and under a light microscope (Biorevo BZ-9000; Keyence, Osaka, Japan). For alizarin red staining, cells were rinsed with PBS three times, fixed in 4% paraformaldehyde phosphate buffer, and stained using an Osteogenesis Assay Kit (ECM815; Millipore) per the manufacturer’s instructions.

Yoshitani J., Kabata T., Arakawa H., Kato Y., Nojima T., Hayashi K., Tokoro M., Sugimoto N., Kajino Y., Inoue D., Ueoka K., Yamamuro Y, & Tsuchiya H. (2020). Combinational therapy with antibiotics and antibiotic-loaded adipose-derived stem cells reduce abscess formation in implant-related infection in rats. Scientific Reports, 10, 11182.

+ Open protocol

+ Expand

Immunocytochemistry Protocol for Studying CY5-3pRNA Stimulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

RAW264.7 cells seeded on a coverslip (Matsunami Glass, Osaka, Japan) were treated with THGP for 24 h, stimulated with Cy5-3pRNA, and incubated at 37 °C for 2 h. Cells were fixed with 4% paraformaldehyde/phosphate buffer (Wako) for 20 min, permeabilized with 0.2% Triton-X for 10 min at room temperature, and blocked with 1% BSA in PBS. The coverslips were incubated for 1 h in the primary antibody diluted in 1% BSA in PBS. Following washing with PBS, the coverslips were incubated for 1 h in appropriate secondary antibodies conjugated with Alexa Fluor 488/594 (Invitrogen) diluted in 1% BSA in PBS. Subsequently, the coverslips were mounted in Slowfade Gold antifade reagent (Invitrogen) with Hoechst 33342 (Invitrogen). Confocal microscopy was performed with an IX-81S confocal microscope (Olympus, Tokyo, Japan). More than 30 cells in each condition were randomly chosen and representative images are shown in Figures.

Baidya S., Nishimoto Y., Sato S., Shimada Y., Sakurai N., Nonaka H., Noguchi K., Kido M., Tadano S., Ishikawa K., Li K., Okubo A., Yamada T., Orba Y., Sasaki M., Sawa H., Miyamoto H., Takada A., Nakamura T, & Takaoka A. (2021). Dual Effect of Organogermanium Compound THGP on RIG-I-Mediated Viral Sensing and Viral Replication during Influenza a Virus Infection. Viruses, 13(9), 1674.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!