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Paraformaldehyde phosphate

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Paraformaldehyde phosphate is a chemical compound used as a fixative in various laboratory applications. It serves as a stabilizing agent, preserving the structural integrity of biological samples. The core function of paraformaldehyde phosphate is to crosslink proteins and nucleic acids, enabling effective preservation and analysis of cellular and tissue specimens.

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Lab products found in correlation

Carboxymethyl cellulose, by Fujifilm (5 mentions) Methylene blue, by Nacalai Tesque (5 mentions)

Spelling variants of this product

4% paraformaldehyde phosphate buffer solution (23 citations) Phosphate-buffered 4% paraformaldehyde (2 citations) 4% paraformaldehyde phosphate buffer (2 citations)

5 protocols using paraformaldehyde phosphate

1

SARS-CoV-2 Plaque Assay in VeroE6/TMPRSS2 Cells

Cited 3 times
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Plaque assay (Figure 4B) was performed as previously described (Kimura et al., 2022b (link); Motozono et al., 2021 (link); Saito et al., 2022 (link); Suzuki et al., 2022 (link); Yamasoba et al., 2022b (link)). Briefly, one day before infection, VeroE6/TMPRSS2 cells (100,000 cells) were seeded into a 24-well plate and infected with SARS-CoV-2 (1, 10, 100 and 1,000 TCID50) at 37°C for 1 hour. Mounting solution containing 3% FBS and 1.5% carboxymethyl cellulose (Wako, Cat# 039-01335) was overlaid, followed by incubation at 37°C. At 3 d.p.i., the culture medium was removed, and the cells were washed with PBS three times and fixed with 4% paraformaldehyde phosphate (Nacalai Tesque, Cat# 09154-85). The fixed cells were washed with tap water, dried, and stained with staining solution [0.1% methylene blue (Nacalai Tesque, Cat# 22412-14) in water] for 30 minutes. The stained cells were washed with tap water and dried, and the size of plaques was measured using Fiji software v2.2.0 (ImageJ).
Kimura I., Yamasoba D., Tamura T., Nao N., Suzuki T., Oda Y., Mitoma S., Ito J., Nasser H., Zahradnik J., Uriu K., Fujita S., Kosugi Y., Wang L., Tsuda M., Kishimoto M., Ito H., Suzuki R., Shimizu R., Begum M.M., Yoshimatsu K., Kimura K.T., Sasaki J., Sasaki-Tabata K., Yamamoto Y., Nagamoto T., Kanamune J., Kobiyama K., Asakura H., Nagashima M., Sadamasu K., Yoshimura K., Shirakawa K., Takaori-Kondo A., Kuramochi J., Schreiber G., Ishii K.J., Hashiguchi T., Ikeda T., Saito A., Fukuhara T., Tanaka S., Matsuno K, & Sato K. (2022). Virological characteristics of the SARS-CoV-2 Omicron BA.2 subvariants, including BA.4 and BA.5. Cell, 185(21), 3992-4007.e16.
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2

SARS-CoV-2 Plaque Assay in VeroE6/TMPRSS2 Cells

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
A plaque assay (Figure 4J) was performed as previously described (Kimura et al., 2022b (link), 2022c (link); Yamasoba et al., 2022a (link); Suzuki et al., 2022 (link)). Briefly, one day before infection, VeroE6/TMPRSS2 cells (100,000 cells) were seeded into a 24-well plate and infected with SARS-CoV-2 (0.5, 5, 50 and 500 TCID50) at 37°C for 1 hour. Mounting solution containing 3% FBS and 1.5% carboxymethyl cellulose (Wako, Cat# 039-01335) was overlaid, followed by incubation at 37°C. At 3 d.p.i., the culture medium was removed, and the cells were washed with PBS three times and fixed with 4% paraformaldehyde phosphate (Nacalai Tesque, Cat# 09154-85). The fixed cells were washed with tap water, dried, and stained with staining solution [0.1% methylene blue (Nacalai Tesque, Cat# 22412-14) in water] for 30 minutes. The stained cells were washed with tap water and dried, and the size of plaques was measured using Fiji software v2.2.0 (ImageJ).
Saito A., Tamura T., Zahradnik J., Deguchi S., Tabata K., Anraku Y., Kimura I., Ito J., Yamasoba D., Nasser H., Toyoda M., Nagata K., Uriu K., Kosugi Y., Fujita S., Shofa M., Monira Begum M., Shimizu R., Oda Y., Suzuki R., Ito H., Nao N., Wang L., Tsuda M., Yoshimatsu K., Kuramochi J., Kita S., Sasaki-Tabata K., Fukuhara H., Maenaka K., Yamamoto Y., Nagamoto T., Asakura H., Nagashima M., Sadamasu K., Yoshimura K., Ueno T., Schreiber G., Takaori-Kondo A., Shirakawa K., Sawa H., Irie T., Hashiguchi T., Takayama K., Matsuno K., Tanaka S., Ikeda T., Fukuhara T, & Sato K. (2022). Virological characteristics of the SARS-CoV-2 Omicron BA.2.75 variant. Cell Host & Microbe, 30(11), 1540-1555.e15.
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3

SARS-CoV-2 Plaque Assay in VeroE6/TMPRSS2 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
The plaque assay was performed as previously described2 ,23 (link). In brief, one day before infection, VeroE6/TMPRSS2 cells (100,000 cells) were seeded into a 24-well plate and infected with SARS-CoV-2 (10,000 TCID50) at 37 °C. At 2 h.p.i., mounting solution containing 3% FBS and 1.5% carboxymethyl cellulose (Wako, 039-01335) was overlaid, followed by incubation at 37 °C. At 3 d.p.i., the culture medium was removed, and the cells were washed with PBS three times and fixed with 4% paraformaldehyde phosphate (Nacalai Tesque, 09154-85). The fixed cells were washed with tap water, dried and stained with staining solution (0.1% methylene blue (Nacalai Tesque, 22412-14) in water) for 30 min. The stained cells were washed with tap water and dried, and the size of plaques was measured using Fiji software v.2.2.0 (ImageJ).
Suzuki R., Yamasoba D., Kimura I., Wang L., Kishimoto M., Ito J., Morioka Y., Nao N., Nasser H., Uriu K., Kosugi Y., Tsuda M., Orba Y., Sasaki M., Shimizu R., Kawabata R., Yoshimatsu K., Asakura H., Nagashima M., Sadamasu K., Yoshimura K., Sawa H., Ikeda T., Irie T., Matsuno K., Tanaka S., Fukuhara T, & Sato K. (2022). Attenuated fusogenicity and pathogenicity of SARS-CoV-2 Omicron variant. Nature, 603(7902), 700-705.
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4

Quantification of SARS-CoV-2 Plaque Formation

Cited 2 times
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Plaque assay (Figure 3D) was performed as previously described (Motozono et al., 2021 (link); Saito et al., 2022 (link); Suzuki et al., 2022 (link)). Briefly, one day before infection, VeroE6/TMPRSS2 cells (100,000 cells) were seeded into a 24-well plate and infected with SARS-CoV-2 (1, 10, 100 and 1,000 TCID50) at 37°C for 2 h. Mounting solution containing 3% FBS and 1.5% carboxymethyl cellulose (Wako, Cat# 039-01335) was overlaid, followed by incubation at 37°C. At 3 d.p.i., the culture medium was removed, and the cells were washed with PBS three times and fixed with 4% paraformaldehyde phosphate (Nacalai Tesque, Cat# 09154-85). The fixed cells were washed with tap water, dried, and stained with staining solution [0.1% methylene blue (Nacalai Tesque, Cat# 22412-14) in water] for 30 m. The stained cells were washed with tap water and dried, and the size of plaques was measured using Fiji software v2.2.0 (ImageJ).
Yamasoba D., Kimura I., Nasser H., Morioka Y., Nao N., Ito J., Uriu K., Tsuda M., Zahradnik J., Shirakawa K., Suzuki R., Kishimoto M., Kosugi Y., Kobiyama K., Hara T., Toyoda M., Tanaka Y.L., Butlertanaka E.P., Shimizu R., Ito H., Wang L., Oda Y., Orba Y., Sasaki M., Nagata K., Yoshimatsu K., Asakura H., Nagashima M., Sadamasu K., Yoshimura K., Kuramochi J., Seki M., Fujiki R., Kaneda A., Shimada T., Nakada T.A., Sakao S., Suzuki T., Ueno T., Takaori-Kondo A., Ishii K.J., Schreiber G., Sawa H., Saito A., Irie T., Tanaka S., Matsuno K., Fukuhara T., Ikeda T, & Sato K. (2022). Virological characteristics of the SARS-CoV-2 Omicron BA.2 spike. Cell, 185(12), 2103-2115.e19.
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5

SARS-CoV-2 Plaque Assay Protocol

Cited 6 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Plaque assay (Fig. 3E) was performed as previously described (2 (link), 3 (link), 6 (link), 14 (link), 15 (link), 24 (link)). Briefly, 1 day before infection, VeroE6/TMPRSS2 cells (100,000 cells) were seeded into a 24-well plate and infected with SARS-CoV-2 (1, 10, 100, and 1,000 TCID50) at 37°C for 1 hour. A mounting solution containing 3% FBS and 1.5% carboxymethyl cellulose (Wako, Cat# 039-01335) was overlaid, followed by incubation at 37°C. At 3 d.p.i., the culture medium was removed, and the cells were washed with PBS three times and fixed with 4% paraformaldehyde phosphate (Nacalai Tesque, Cat# 09154-85). The fixed cells were washed with tap water, dried, and stained with a staining solution [0.1% methylene blue (Nacalai Tesque, Cat# 22412-14) in water] for 30 min. The stained cells were washed with tap water and dried, and the size of the plaques was measured using Fiji software v2.2.0 (ImageJ).
Kimura I., Yamasoba D., Nasser H., Ito H., Zahradnik J., Wu J., Fujita S., Uriu K., Sasaki J., Tamura T., Suzuki R., Deguchi S., Plianchaisuk A., Yoshimatsu K., Kazuma Y., Mitoma S., Schreiber G., Asakura H., Nagashima M., Sadamasu K., Yoshimura K., Takaori-Kondo A., Ito J., Shirakawa K., Takayama K., Irie T., Hashiguchi T., Nakagawa S., Fukuhara T., Saito A., Ikeda T, & Sato K. (2023). Multiple mutations of SARS-CoV-2 Omicron BA.2 variant orchestrate its virological characteristics. Journal of Virology, 97(10), e01011-23.
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