Phosphate buffer

Manufactured by Nacalai Tesque

Phosphate buffer is a type of buffer solution commonly used in laboratory settings. It maintains a stable pH range, typically between 6.0 and 8.0, which is suitable for various biological and chemical applications. The core function of phosphate buffer is to help maintain a desired pH level in experimental conditions.

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Lab products found in correlation

Balb c mice, by Japan SLC (1 mentions) Pneumovax np, by MSD (1 mentions) Provis ax 80 microscope, by Olympus (1 mentions) Female balb c mice, by Japan SLC (1 mentions) Mplas, by InvivoGen (1 mentions) Fv1000d, by Olympus (1 mentions) Paraformaldehyde (pfa), by Nacalai Tesque (1 mentions) A1rmp microscope, by Nikon (1 mentions) Bz x710, by Keyence (1 mentions)

6 protocols using phosphate buffer

Mucosal Vaccine Composition Evaluation

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Example 5

To 87 μL (1150 μg/mL) of a pneumococcal capsular polysaccharide-containing solution (PNEUMOVAX NP, MSD K.K.), and 2.5 μL (2 mg/mL) of a solution of a lipopolysaccharide derived from Pantoea agglomerans (available from Nacalai Tesque), a phosphate buffer (available from Nacalai Tesque) was added to prepare 100 μL of a mucosal vaccine composition.

Four mice (female BALB/C mice aged 8 weeks, Japan SLC, Inc.) were anesthetized, and 20 μL of the prepared mucosal vaccine composition was sublingually administered to each mouse. After one week from the administration, the mice were anesthetized again, and 20 μL of the prepared mucosal vaccine composition was sublingually administered to each mouse. After one week from the second administration, a serum and a nasal cavity washing liquid of each mouse were collected, and a pneumococcal capsular polysaccharide-specific IgG titer in a serum and a pneumococcal capsular polysaccharide-specific IgA titer in a nasal cavity washing liquid were determined by the ELISA method. Specific determination methods will be described later.

US10092642B2. Mucosal vaccine composition (2018-10-09). NITTO DENKO CORPORATION [JP]. Inventors: Masahiro Fukasaka [JP], Mitsuhiko Hori [JP], Katsuyuki Okubo [JP], Daisuke Asari [JP], Arimichi Okazaki [JP], Eiji Kiyotoh [JP], Kyohei Matsushita [JP].

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Tissue Fixation and Staining

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Mouse tissues were fixed in 10% formalin in phosphate buffer (Nakarai Tesque) and then embedded in paraffin. Hematoxylin and eosin staining was performed according to standard procedures. Images were captured using a Provis AX80 microscope (Olympus) equipped an OLYMPUS DP70 digital camera, and detected using a DP manager system (Olympus).

Mitagami Y., Yasunaga J.I., Kinosada H., Ohshima K, & Matsuoka M. (2015). Interferon-γ Promotes Inflammation and Development of T-Cell Lymphoma in HTLV-1 bZIP Factor Transgenic Mice. PLoS Pathogens, 11(8), e1005120.

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Mucosal Vaccination for HPV16 Immunity

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Example 6

To 61 μL (820 μg/mL) of an HPV16 recombinant protein-containing solution (HPV16, available from PROSPEC), and 2.5 μL (2 mg/mL) of a solution of a lipopolysaccharide derived from Pantoea agglomerans (available from Nacalai Tesque), a phosphate buffer (available from Nacalai Tesque) was added to prepare 100 μL of a mucosal vaccine composition.

Four mice (female BALB/C mice aged 8 weeks, Japan SLC, Inc.) were anesthetized, and 20 μL of the prepared mucosal vaccine composition was sublingually administered to each mouse. After one week from the administration, the mice were anesthetized again, and 20 μL of the prepared mucosal vaccine composition was sublingually administered to each mouse. After one week from the second administration, a serum and a nasal cavity washing liquid of each mouse were collected, and an HPV16-specific IgG titer in a serum and an HPV16-specific IgA titer in a nasal cavity washing liquid were determined by the ELISA method. Specific determination methods will be described later.

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Rotavirus Vaccine Composition and Evaluation

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Example 14

To 200 μL of an attenuated live rotavirus-containing solution (RotaTeq mixture for internal use, available from MSD K.K.), 50 μL (2 mg/mL) in Example 11, 5 μL in Example 12, or 0.5 μL in Example 13 of a solution of a lipopolysaccharide derived from Pantoea agglomerans (available from Nacalai Tesque), or 5 μL of a solution of a glucopyranosyl lipid (MPLAs, available from InvivoGen) (2 mg/mL) in Comparative Example 14 was added, and a phosphate buffer (available from Nacalai Tesque) was added to prepare 1000 μL of a vaccine composition.

Six mice (female BALB/C mice aged 8 weeks, Japan SLC, Inc.) are anesthetized, and 100 μL of the prepared vaccine composition is subcutaneously administered to each mouse. After one week from the administration, the mice are anesthetized again, and 100 μL of the prepared vaccine composition is subcutaneously administered to each mouse. After one week from the second administration, a mouse serum is collected, and an antigen-specific IgG titer in the serum is determined by the ELISA method.

US09962439B2. Injectable vaccine composition (2018-05-08). NITTO DENKO CORPORATION [JP]. Inventors: Kyohei Matsushita [JP], Masahiro Fukasaka [JP], Arimichi Okazaki [JP], Eiji Kiyotoh [JP], Katsuyuki Okubo [JP], Daisuke Asari [JP], Mitsuhiko Hori [JP].

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Immunofluorescence Analysis of NF-κB p65

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Peritoneal macrophages cultured on glass coverslips in a 6-well plate were washed once with ice-cold PBS and fixed with 4% paraformaldehyde in phosphate buffer (Nacalai Tesque) for 15 min at room temperature (RT, 20–26°C). Fixed cells were washed three times with PBS and permeabilized with 0.5% Triton X-100 in PBS for 15 min at RT. Subsequently, cells were washed three times with PBS and blocked with 2% BSA in PBS for 1 h at RT. The cells were stained with the anti-NF-κB p65 antibody listed in Supplementary Table 2, and the cell nuclei were stained with DAPI. The cells on glass coverslips were analyzed by confocal fluorescent microscopy (FV1000-D, Olympus, Tokyo, Japan).

Kawano M., Miyoshi M, & Miyazaki T. (2019). Lactobacillus helveticus SBT2171 Induces A20 Expression via Toll-Like Receptor 2 Signaling and Inhibits the Lipopolysaccharide-Induced Activation of Nuclear Factor-kappa B and Mitogen-Activated Protein Kinases in Peritoneal Macrophages. Frontiers in Immunology, 10, 845.

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Spheroid Diameter, Viability, and Imaging

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The diameter of the spheroids was measured using a microscope with an image analysis system (BZ-X710, KEYENCE, Osaka, Japan). Twenty spheroids were collected using a micropipette and were dispersed using 0.25% (w/v) trypsin-EDTA. After staining with trypan blue solution, the number of live cells was measured by counting the unstained cells using microscopy, and the number of live cells in each spheroid was calculated. The cell viability in spheroids could not be calculated because dead cells were hardly liberated from cell debris after dispersion of the spheroids.
Live/Dead Imaging The spheroids were stained with 1 µM CFSE and 5 µg/mL propidium iodide in Opti-MEM (30 min, 37 °C). After washing three times with phosphatebuffered saline (PBS), the spheroids were fixed with 4% paraformaldehyde in phosphate buffer (Nacalai Tesque, Inc.) overnight at 4 °C. After fixing, the spheroids were optically cleared using the ScaleS method. 21) (link) In brief, the spheroids were incubated with ScaleSQ(0) (22.5% (w/v) D[-]-sorbitol and 9.1 M urea [pH 8.4]) for 24 h at 4 °C and then with ScaleS4(0) (40% (w/v) D[-]-sorbitol, 10% (w/v) glycerol, 4 M urea, and 20% (v/v) dimethyl sulfoxide) for 24 h at 4 °C. The optically cleared spheroids were observed using confocal fluorescence microscopy (A1R MP microscope, Nikon, Tokyo, Japan).

Mizukami Y., Takahashi Y., Shimizu K., Konishi S., Takakura Y, & Nishikawa M. (2021). Calcium Peroxide-Containing Polydimethylsiloxane-Based Microwells for Inhibiting Cell Death in Spheroids through Improved Oxygen Supply. Biological & pharmaceutical bulletin, 44(10).

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