Paraformaldehyde (pfa)

Manufactured by Keygen Biotech

Sourced in China

Paraformaldehyde is a solid polymer of formaldehyde. It is a widely used chemical compound in laboratory settings, primarily as a fixative and preservative for biological samples.

Automatically generated - may contain errors

Lab products found in correlation

Matrigel, by BD (5 mentions) 4 6 diamidino 2 phenylindole (dapi), by Keygen Biotech (3 mentions) Tdt enzyme reaction solution, by Keygen Biotech (2 mentions) Fluorescence microscope, by Olympus (2 mentions) Crystal violet, by Merck Group (2 mentions) Jem 1400plus, by JEOL (2 mentions) Glutaraldehyde, by Wuhan Servicebio Technology (2 mentions) Dm60008, by Leica (2 mentions) Crystal violet staining solution, by Keygen Biotech (2 mentions) Tunel kit, by Keygen Biotech (1 mentions) Tdt mediated dutp nick end labeling tunel kit, by Keygen Biotech (1 mentions) Bovine serum albumin (bsa), by GE Healthcare (1 mentions) Hoechst 33342, by Keygen Biotech (1 mentions) Transwell chamber, by Corning (1 mentions) Triton x 100, by Merck Group (1 mentions) Diamidino phenyl indole dapi, by Merck Group (1 mentions) Anti human serum afp antibody, by Santa Cruz Biotechnology (1 mentions) Anti human serum alb antibody, by Santa Cruz Biotechnology (1 mentions) Confocal microscope, by Zeiss (1 mentions) Bovine serum albumin (bsa), by Keygen Biotech (1 mentions)

20 protocols using paraformaldehyde (pfa)

Cell Viability and Apoptosis Assessment

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Cells were seeded into 96‐well plates at a density of 1 × 10⁴ cells per well and were treated with drugs the following day. After treatment for the noted time, 20 μl of CCK8 reagent (Vicmed) was added to each well, the plate was incubated in the dark for 30 min, and the absorbance at 450 nm was detected by a microplate reader.
Apoptosis was measured via a TUNEL kit (KeyGen), according to the manufacturer's instructions. After treatment, the cells were fixed with 4% paraformaldehyde (KeyGen) for 30 min, and the cells were then permeabilized for 5 min. TDT enzyme reaction solution (KeyGen) was added to each well, and reactions were incubated in the dark at 37°C for 1 h. The streptavidin TRITC labeling solution (KeyGen) was added, and the mixture was incubated in the dark in a humidified environment at 37°C for 30 min. After washing three times with PBS, DAPI was added for 10 min to counterstain nuclei. The results were observed with a fluorescence microscope.

Cheng B., Hong X., Wang L., Cao Y., Qin D., Zhou H, & Gao D. (2022). Curzerene suppresses progression of human glioblastoma through inhibition of glutathione S‐transferase A4. CNS Neuroscience & Therapeutics, 28(5), 690-702.

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Quantification of Apoptosis via TUNEL Assay

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Apoptosis was measured via a TdT-mediated dUTP Nick-End Labeling (TUNEL) kit (KeyGen, Jiangsu, China). Briefly, the treated cells were fixed with 4% paraformaldehyde (KeyGen, Jiangsu, China) for 30 min and permeabilized for 5 min. TDT enzyme reaction solution (KeyGen, Jiangsu, China) was added to each well, and reactions were incubated in the dark at room temperature for 1 h. The cells were mixed with streptavidin labeling solution (KeyGen, Jiangsu, China) and incubated in the dark for 30 min. DAPI was added to counterstain the nuclei after washing the cells three times with PBS. After staining, the percentage of TUNEL-positive cells was counted.

Cheng B., Wang Y., Ayanlaja A.A., Zhu J., Kambey P.A., Qiu Z., Zhang C, & Hu W. (2022). Glutathione S-Transferases S1, Z1 and A1 Serve as Prognostic Factors in Glioblastoma and Promote Drug Resistance through Antioxidant Pathways. Cells, 11(20), 3232.

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Interleukin-4 Stimulation of Transfected DU145 Cells

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Transfected DU145 cells were seeded on coverslips, stimulated with 50 ng/ml human interleukin (IL)-4 (GE Healthcare Life Sciences) for 30 min, fixed with 4% paraformaldehyde (KeyGen Biotech. Co. Ltd.), blocked with 3% bovine serum albumin (BSA; GE Healthcare Life Sciences) for 1 h, and incubated with primary antibodies overnight at 4°C. Fluorescein isothiocyanate-labeled secondary antibody was then added for 2 h at 37°C, and following staining with DAPI (KeyGen Biotech. Co. Ltd.) for 5 min, the cells were subjected to confocal laser scanning (Nikon Corporation, Tokyo, Japan).

WANG N., TAO L., ZHONG H., ZHAO S., YU Y., YU B., CHEN X., GAO J, & WANG R. (2015). miR-135b inhibits tumour metastasis in prostate cancer by targeting STAT6. Oncology Letters, 11(1), 543-550.

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Apoptosis Assay for IBV Infection

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HD11 cells were pre-incubated in 96-well plates and infected with IBV at 10 MOI. To assess apoptosis, the condensed and fragmented nuclei were observed using Hoechst 33342 staining (KeyGEN Biotech, Nanjing, Jiangsu Province, China). At the specified time points, the cells were immobilized with 4% paraformaldehyde (KeyGEN Biotech, Nanjing, Jiangsu Province, China) for 30 min and then incubated with Hoechst 33342 (KeyGEN Biotech) in the dark for 15 min. The typical apoptotic morphological changes were observed using a fluorescence microscope (Olympus IX71, Olympus, Tokyo, Japan) with UV excitation at 350 nm.

Han X., Tian Y., Guan R., Gao W., Yang X., Zhou L, & Wang H. (2017). Infectious Bronchitis Virus Infection Induces Apoptosis during Replication in Chicken Macrophage HD11 Cells. Viruses, 9(8), 198.

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Transwell Assay for Cell Migration and Invasion

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Cells were seeded into Transwell chambers (Corning, USA) in triplicate at a density of 5 × 10⁴ cells per well in 200 μl of serum-free culture medium. For cell invasion assays, the Transwell membranes were precoated with Matrigel (BD, USA) before cell seeding. Then, 600 μl of culture medium supplemented with 20% FBS was added into the lower compartment of the Transwell chambers as a chemoattractant. The cells were allowed to migrate for 24 h or invade for 48 h and were then fixed with 4% paraformaldehyde (KeyGEN BioTECH) for 30 min and stained with crystal violet staining solution for 10 min. Crystal violet-stained cells adhering to the lower surface of the Transwell membranes were counted under a microscope in five random fields. Data were analyzed using ImageJ software.

Jiang K., Dong C., Yin Z., Li R., Mao J., Wang C., Zhang J., Gao Z., Liang R., Wang Q, & Wang L. (2020). Exosome-derived ENO1 regulates integrin α6β4 expression and promotes hepatocellular carcinoma growth and metastasis. Cell Death & Disease, 11(11), 972.

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Immunofluorescence Analysis of Hepatic Markers

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Cells were washed twice with cold PBS and fixed in 4% paraformaldehyde (KeyGen Biotech Co., Ltd.) in PBS for 30 min and permeabilized with PBS containing 0.1% Triton X-100 (Sigma-Aldrich; Merck KGaA) for 20 min. The samples were incubated with anti-human serum AFP antibody (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), anti-human serum ALB antibody (Santa Cruz Biotechnology, Inc.), and anti-human serum cytochrome P450 3A4 (CYP3A4) antibody (Santa Cruz Biotechnology, Inc.), followed by incubation with second antibody conjugated with fluorescent phycobilioroteins, Dylight 594 and Alexa 488 goat anti-mouse immunoglobulin G (1:2,000; Sigma-Aldrich; Merck KGaA). Subsequently, the cells were stained with diamidinopheny-lindole (DAPI; Sigma-Aldrich; Merck KGaA) and observed under a fluorescence microscope (Olympus, Tokyo, Japan).

Yu Y.B., Song Y., Chen Y., Zhang F, & Qi F.Z. (2018). Differentiation of umbilical cord mesenchymal stem cells into hepatocytes in comparison with bone marrow mesenchymal stem cells. Molecular Medicine Reports, 18(2), 2009-2016.

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Quantifying Macrophage Phenotypes via Immunofluorescence

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RAW264.7 macrophages seeded on glass-bottom dishes were fixed in 4% paraformaldehyde (KeyGen Biotech, China) and then permeabilized with ice-cold 0.5% TritonX-100. The cells were blocked in PBS containing 10% bovine serum albumin (KeyGen Biotech, China) for 30 min and then were incubated with rabbit monoclonal antibody inducible nitric oxide synthase (iNOS) (Bioss Antibodies, China) and rat monoclonal antibody F4/80 (Abcam, USA) overnight at 4°C. After washing with PBS, cells were incubated with Alexa Fluor 488 goat anti-rabbit IgG (Abcam, USA) and Alexa Fluor 647 goat anti-rat IgG (Abcam, USA) for 1 h at 37°C. Finally, cells were counterstained with 50 mg/ml 4′,6-diamidino-2-phenylindole (DAPI) (KeyGen Biotech, China) before capturing images with a confocal microscope (Zeiss, Germany).

Wang Q., Li S., Tang X., Liang L., Wang F, & Du H. (2019). Lipocalin 2 Protects Against Escherichia coli Infection by Modulating Neutrophil and Macrophage Function. Frontiers in Immunology, 10, 2594.

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Cell Migration and Invasion Assay

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The 24-transwell migration chambers (8 μm pore) (Cat#3464, Corning Life Sciences, USA) were used to assess migration and invasion ability, coated with BD Matrigel^TM Basement Membrane Matrix (Cat#354234, BD Biosciences, USA) (1:1 dilution, 50 μl/cm2) or not. The serum-free medium was in the upper wells while the medium with 20% FBS was in the bottom wells. Ten thousand cells were incubated in the upper wells for 48 h, then transwell chambers were fixed by 4% paraformaldehyde (Cat#KGIHC007, KeyGEN BioTECH, China) and stained with crystal violet (Cat#94448, Sigma, USA).

Wang Y., Li Z., Xu S., Li W., Chen M., Jiang M, & Fan X. (2022). LncRNA FIRRE functions as a tumor promoter by interaction with PTBP1 to stabilize BECN1 mRNA and facilitate autophagy. Cell Death & Disease, 13(2), 98.

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Transwell Invasion and Migration Assay

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BD Transwell chambers were coated with Matrigel for the Transwell invasion assay and without Matrigel for the migration assay (BD Biosciences, Franklin Lakes, NJ, USA). HT-29 and HCT116 cells were treated with 5 and 10 μM DMC-BH for 12 h. Then, a total of 1×10⁵ cells per chamber were added to the top chamber with serum-free DMEM, while DMEM with 10% FBS was added to the bottom chamber at 37° C and 5% CO₂. After 24 h of incubation, the cells on the inner surface of the upper chamber were removed. Then, the invaded cells on the outer surface were fixed with 4% paraformaldehyde (KeyGEN BioTECH, Nanjing, China) and stained with 0.1% crystal violet solution (Sigma-Aldrich, St. Louis, MO, USA) for 5 min. The invading cells were counted in three independent fields (100×) under a microscope.

Liu G., Chen J, & Bao Z. (2023). Promising antitumor effects of the curcumin analog DMC-BH on colorectal cancer cells. Aging (Albany NY), 15(6), 2221-2236.

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Invasion Assay for Breast Cancer Cells

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For the invasion assay, polycarbonate filters were coated with 20 μL of Matrigel (1:5; BD Bioscience) and placed in a Transwell Permeable chamber (Costar, USA). MCF-10A and MCF-7 cells were plated into 96-well plates at 4×10⁴ cells/mL for each well for the invasion assay; 1×10⁵ cells were counted and suspended in 100 µL of serum-free medium and then were added to the Transwell upper chamber (Costar, Corning, NY, USA) after transfection; Next, 600 µL of complete medium (DMEM; Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) was added into the lower chamber. Following a 48-h incubation, the untransferred cells and Matrigel were removed using a cotton swab. The transfected cells were fixed using 4% paraformaldehyde for 30 min, permeabilized with 0.2% Triton X-100 (KeyGen, Nanjing, China) for 15 min, and stained using 0.05% gentian violet (KeyGen) dye for 5 min. The cells in five random regions were counted, and the average count was calculated. The experiment was performed in triplicate.

Tao Z., Suo H., Zhang L., Jin Z., Wang Z., Wang D., Wu M., Peng N., Zhao Y, & Chen B. (2020). MRPL13 is a Prognostic Cancer Biomarker and Correlates with Immune Infiltrates in Breast Cancer. OncoTargets and therapy, 13, 12255-12268.

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