Log In Sign up
The largest database of trusted experimental protocols

Phorbol 12 myristate 13 acetate pma

Manufactured by Cayman Chemical
Visit Supplier
Sourced in United States

Phorbol 12-myristate 13-acetate (PMA) is a chemical compound that acts as a potent activator of protein kinase C (PKC). It is a white crystalline solid and is commonly used as a research tool in cell biology and biochemistry experiments.

Automatically generated - may contain errors

Lab products found in correlation

Ionomycin, by Cayman Chemical (3 mentions) Erythrocyte depletion, by Miltenyi Biotec (2 mentions) Easysep mouse neutrophil enrichment kit, by STEMCELL (2 mentions) Macsxpress whole blood neutrophil isolation kit, by Miltenyi Biotec (2 mentions) Arginylglycylaspartic acid, by Merck Group (1 mentions) Phytohemagglutinin p pha, by Merck Group (1 mentions) Anti cd274, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Welgene (1 mentions) Anti cd11b, by BD (1 mentions) Rpmi 1640 medium without phenol red, by Thermo Fisher Scientific (1 mentions) Heparin natrium 25000, by Ratiopharm (1 mentions) Klebsiella pneumoniae lipopolysaccharide lps, by Merck Group (1 mentions) Dnase 1, by Roche (1 mentions) Ham s f 12 nutrient mix, by Thermo Fisher Scientific (1 mentions) Luminol, by Merck Group (1 mentions) Hybri max, by Merck Group (1 mentions) Acetylcholine chloride, by Merck Group (1 mentions) Wortmannin, by Cayman Chemical (1 mentions) Bapta am, by Thermo Fisher Scientific (1 mentions) Nω nitro l arginine methyl ester hydrochloride l name, by Merck Group (1 mentions)

13 protocols using phorbol 12 myristate 13 acetate pma

1

Comparative Evaluation of Surfactant Preparations

Check if the same lab product or an alternative is used in the 5 most similar protocols
Two natural surfactant preparations common in clinical use were examined: Curosurf® (Chiesi, Hamburg, Germany) is prepared from minced porcine lungs and supplied as a ready-to-use intratracheal suspension. Alveofact® (Lyomark Pharma, Oberhaching, Germany) is a surfactant isolated from bovine lung lavage and prepared as a lyophilized powder that has to be reconstituted with the provided solvent prior to use. The surfactants were used and stored according to the manufacturers’ recommendations. To obtain approximately similar concentrations in vitro we covered a range of concentrations from 0.001–2.5 mg/ml.
Moreover, phorbol-12-myristate-13-acetate (PMA), a common chemical stimulator for NETosis (10 (link)), from Cayman Chemical (USA) was used.
Schulz A., Pagerols Raluy L., Kolman J.P., Königs I., Trochimiuk M., Appl B., Reinshagen K., Boettcher M, & Trah J. (2021). The Inhibitory Effect of Curosurf® and Alveofact® on the Formation of Neutrophil Extracellular Traps. Frontiers in Immunology, 11, 582895.
+ Open protocol
+ Expand
2

Immune Cell Activation and Proliferation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The following reagents were obtained from the respective suppliers: phorbol-12-myristate 13-acetate (PMA; Cayman Chemical (Ann Arbor, MI, USA)); 1,25-dihydroxy vitamin D3 (Toronto Research Chemicals, Inc., Toronto, ON, Canada); Roswell Park Memorial Institute (RPMI)-1640, Dulbecco modified Eagle medium (DMEM), Dulbecco phosphate-buffered saline (DPBS), and fetal bovine serum (FBS) (Welgene, Inc., Daegu, Republic of Korea); Interferon-gamma (IFN-γ) and anti-MER-TK antibodies (R&D Systems, Minneapolis, MN, USA); Arginylglycylaspartic acid (RGD), phytohemagglutinin P (PHA), and LPS from Escherichia coli K12 (Sigma-Aldrich, St. Louis, MO, USA); carboxyfluorescein succinimidyl ester (CFSE) CellTrace™ Cell Proliferation Kit, and antibiotic-antimycotic (Invitrogen; Thermo Fisher Scientific Inc., Waltham, MA, USA); ethylenediaminetetraacetic acid (EDTA; Amresco, Inc., Solon, OH, USA); Galα1-3Galβ1-4GlcNAcβ-PAA trisaccharide (α-Gal; GlycoTech, Gaithersburg, MD, USA); anti-CD14, anti-CD274, and anti-CD80 (eBioscience Inc. (San Diego, CA, USA); anti-CD16 (Biolegend Inc., San Diego, CA, USA); anti-DHRS9 (Abnova, Taiwan, China); anti-CD11b (BD Biosciences Inc., Franklin Lakes, NJ, USA); anti-CD209 (Serotec Bio-Rad Inc., CA, USA); and FITC-conjugated, in addition to other antibodies (Santa Cruz Biotechnology Inc., Dallas, TX, USA).
Pham H.L., Hoang T.X, & Kim J.Y. (2023). Human Regulatory Macrophages Derived from THP-1 Cells Using Arginylglycylaspartic Acid and Vitamin D3. Biomedicines, 11(6), 1740.
+ Open protocol
+ Expand
3

Healthy Donor Whole Blood Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
We purchased deidentified, healthy human donor whole blood samples from a commercial vendor (Research Blood Components, Watertown, MA, USA). Protocols were approved by the institutional review board at Massachusetts General Hospital (MGH). The blood was collected in sodium heparin precoated vacuum tubes (Becton, Dickinson and Company, Franklin Lakes, NJ, USA). Blood samples were transported at room temperature (20 – 24 °C) and delivered to the lab within 6 hours after the blood draw. Aliquots of blood were transferred to small vials and stored at 37°C before running the assay. Some of the samples were stimulated with Phorbol 12-myristate 13-acetate (PMA, Cayman Chemical, Ann Arbor, MI, USA), at 1 or 10 μg/mL, before the assay.
Sakuma M., Wang X., Ellett F., Edd J., Kehinde B., Viens A., Mansour M.K, & Irimia D. (2022). Microfluidic capture of chromatin fibres measures neutrophil extracellular traps (NETs) released in a drop of human blood. Lab on a chip, 22(5), 936-944.
+ Open protocol
+ Expand
4

Murine and Human Neutrophil Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Murine neutrophils were isolated from 8–12 week old C56BL/6 mice. Mice were euthanized by cervical dislocation, and bone marrow cells were harvested from femur and tibia within 30 min. Cell suspensions were passed through 70 µm cell strainer. Neutrophils were prepared using a negative selection kit (EasySep Mouse Neutrophil Enrichment Kit, Stem Cell Technologies), following manufacturer instruction. Cells were cultured in RPMI 1640 media supplemented with 10% heat-inactivated FBS, 100 U/ml penicillin, 100 µg/ml streptomycin, 5 mM HEPES, and 2 mM EDTA. To stimulate neutrophils, 100 nM phorbol 12-myristate 13-acetate (PMA) (Cayman Chemical) was added to the media.
Human neutrophils were isolated from 8 ml of blood collected from healthy donors. Neutrophils were purified using the MACSxpress Whole Blood Neutrophil Isolation Kit (Miltenyi Biotec 130–104-434) followed by erythrocyte depletion (Miltenyi Biotec 130–098-196) according to manufacturer’s instructions.
Ghergurovich J.M., García-Cañaveras J.C., Wang J., Schmidt E., Zhang Z., TeSlaa T., Patel H., Chen L., Britt E.C., Piqueras-Nebot M., Gomez-Cabrera M.C., Lahoz A., Fan J., Beier U.H., Kim H, & Rabinowitz J.D. (2020). A small molecule G6PD inhibitor reveals immune dependence on pentose phosphate pathway. Nature chemical biology, 16(7), 731-739.
+ Open protocol
+ Expand
5

Immune Cell Activation and Oxidative Stress

Check if the same lab product or an alternative is used in the 5 most similar protocols
SYTOX™ Green Nucleic Acid Stain - 5 mM Solution in DMSO (ThermoFisher Scientific, S7020), Molecular Probes™ H2DCFDA (ThermoFisher, Scientific D399), Ionomycin (calcium salt) (Cayman Chemical, 11932), Klebsiella pneumoniae lipopolysaccharide (LPS) (Sigma-Aldrich, L4268), Phorbol 12-myristate 13-acetate (PMA) (Cayman Chemical, 10008014), RPMI 1640 Medium without phenol red (ThermoFisher Scientific, 11835063), Lymphoflot Ficoll-Diatrizoate density gradient solution (Bio Rad, 824012), DNase1 (Roche, 11284932001), Heparin-Natrium-25000-ratiopharm® (Ratiopharm).
Leppkes M., Knopf J., Naschberger E., Lindemann A., Singh J., Herrmann I., Stürzl M., Staats L., Mahajan A., Schauer C., Kremer A.N., Völkl S., Amann K., Evert K., Falkeis C., Wehrfritz A., Rieker R.J., Hartmann A., Kremer A.E., Neurath M.F., Muñoz L.E., Schett G, & Herrmann M. (2020). Vascular occlusion by neutrophil extracellular traps in COVID-19. EBioMedicine, 58, 102925.
+ Open protocol
+ Expand
6

Sourcing of Chemical Reagents

Check if the same lab product or an alternative is used in the 5 most similar protocols
GSK2981278, SR2211, SR0987, phorbol 12-myristate 13-acetate (PMA), and ionomycin were purchased from Cayman Chemicals (Ann Arbor, MI, USA), benzophenone-9 and benzophenone-10 from Carbosynth (Compton, UK), and the remaining benzophenone derivatives, 3-benzylidene camphor (3-BC) and 4-methylbenzylidene camphor (4-MBC) from Merck AG (Glattbrugg, Switzerland). All other chemicals were obtained from Sigma-Aldrich (Buchs, Switzerland). RPMI-1640 cell culture medium (R8758) was purchased from Sigma-Aldrich, Ham’s F12 Nutrient Mix (21765–029) from Gibco (Thermo Fischer Scientific, Waltham, MA, USA), fetal bovine serum (FBS, S1810–500) from Biowest (Nuaillé, France) and defined FBS for the use in Tet-on systems (HyClone, SH30070) from Cytiva (Marlborough, MA, USA).
Inderbinen S.G., Kley M., Zogg M., Sellner M., Fischer A., Kędzierski J., Boudon S., Jetten A.M., Smieško M, & Odermatt A. (2022). Activation of retinoic acid-related orphan receptor γ(t) by parabens and benzophenone UV-filters. Toxicology, 471, 153159.
+ Open protocol
+ Expand
7

Murine and Human Neutrophil Isolation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Murine neutrophils were isolated from 8–12 week old C56BL/6 mice. Mice were euthanized by cervical dislocation, and bone marrow cells were harvested from femur and tibia within 30 min. Cell suspensions were passed through 70 µm cell strainer. Neutrophils were prepared using a negative selection kit (EasySep Mouse Neutrophil Enrichment Kit, Stem Cell Technologies), following manufacturer instruction. Cells were cultured in RPMI 1640 media supplemented with 10% heat-inactivated FBS, 100 U/ml penicillin, 100 µg/ml streptomycin, 5 mM HEPES, and 2 mM EDTA. To stimulate neutrophils, 100 nM phorbol 12-myristate 13-acetate (PMA) (Cayman Chemical) was added to the media.
Human neutrophils were isolated from 8 ml of blood collected from healthy donors. Neutrophils were purified using the MACSxpress Whole Blood Neutrophil Isolation Kit (Miltenyi Biotec 130–104-434) followed by erythrocyte depletion (Miltenyi Biotec 130–098-196) according to manufacturer’s instructions.
Ghergurovich J.M., García-Cañaveras J.C., Wang J., Schmidt E., Zhang Z., TeSlaa T., Patel H., Chen L., Britt E.C., Piqueras-Nebot M., Gomez-Cabrera M.C., Lahoz A., Fan J., Beier U.H., Kim H, & Rabinowitz J.D. (2020). A small molecule G6PD inhibitor reveals immune dependence on pentose phosphate pathway. Nature chemical biology, 16(7), 731-739.
+ Open protocol
+ Expand
8

Quantifying Reactive Oxygen Species

Check if the same lab product or an alternative is used in the 5 most similar protocols
A total of 1 × 105 cells were seeded in 100 µL of medium in white 96-well plates for 48 h. On day 2, the medium was removed. Fifty microlitres (50 μL) Hank’s buffered salt solution (HBSS) for 30 min was added to each well before luminescence measurement. Luminol buffer (10 mg/mL Luminol (Sigma-Aldrich), and HRP 100 U/mL (Sigma-Aldrich) in HBSS) were prepared with and without 60 ng/mL of phorbol 12-myristate 13-acetate (PMA) (Cayman Chemical, Ann Arbor, MI, USA). Fifty microlitres (50 μL) Luminol buffer was added to the cells and the luminescence measurement was started as soon as possible upon the addition because of the immediate reaction. The bioluminescence was measured over a time period of around 2 h. To quantify ROS production, the area under the curve was calculated with Graph Pad Prism software (San Diego, CA, USA).
Sultanli S., Ghumnani S., Ashma R, & Kubatzky K.F. (2021). Plumbagin, a Biomolecule with (Anti)Osteoclastic Properties. International Journal of Molecular Sciences, 22(5), 2779.
+ Open protocol
+ Expand
9

TBEV Neudoerfl E Protein Peptide Synthesis

Check if the same lab product or an alternative is used in the 5 most similar protocols
15-mer peptides overlapped by 11 amino acids spanning the whole TBEV Neudoerfl E protein (UniProtKB: P14336) were synthesized (≥75% purity, GenScript Biotech Corp, Piscataway Township, New Jersey, USA). Lyophilized peptides were resolved in DMSO (Hybri-Max™, Sigma-Aldrich, St. Louis, Missouri, USA), and two peptide pools with 10 µg/ml of each peptide were generated (E1-255: 61 peptides, E245-496: 60 peptides). For ex vivo restimulation, spleen cells were incubated overnight at 37°C/5% CO2 with 1 µg/ml of the respective peptide pool, live MVA (MOI 3), DMSO/R10F (negative control), or a mixture of 30 ng/ml of phorbol 12-myristate 13-acetate (PMA; Cayman Chemical, Ann Arbor, Michigan, USA) and 0.5 µg/ml of ionomycin (Cayman Chemical, Ann Arbor, Michigan, USA) (positive control).
Kubinski M., Beicht J., Zdora I., Biermann J., Puff C., Gerlach T., Tscherne A., Baumgärtner W., Osterhaus A.D., Sutter G., Prajeeth C.K, & Rimmelzwaan G.F. (2023). A recombinant Modified Vaccinia virus Ankara expressing prME of tick-borne encephalitis virus affords mice full protection against TBEV infection. Frontiers in Immunology, 14, 1182963.
+ Open protocol
+ Expand
10

Calcium Signaling Modulators Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
TRPV4 specific agonist GSK1016790A (GSK101, #G0798) and inhibitor GSK2193874 (GSK874, #5106) were obtained from Sigma-Aldrich (St. Louis, MO) and Tocris Bioscience (Bristol, UK), respectively. Compound C (AMPK inhibitor, #11967), STO-609 (calmodulin-dependent kinase kinase (CaMKK) inhibitor, #15325), Wortmannin (PI3K inhibitor, #10010591), LY294002 (Akt inhibitor, #70920), U0126 (ERK1/2 inhibitor, #70970), AICAR (#10010241), Forskolin (#11018), and Phorbol 12-myristate 13-acetate (PMA, #10008014) were obtained from Cayman Chemical (Ann Arbor, MI). Ruthenium Red (Ca2+ fluxes blocker, #557450), and H-89 (PKA inhibitor, #371963) were obtained from Calbiochem (San Diego, CA). Calcium chelator BAPTA/AM (#B-1205) was from Invitrogen (Grand Island, NY). Recombinant mouse TNF-α was from Roche (#11271156001, Indianapolis, IN). Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME, #N5751), Sodium nitroprusside (SNP, #PHR1423), Acetylcholine chloride (#A2661), (R)-(−)-Phenylephrine hydrochloride (PE, #P8155) and other chemicals were obtained from Sigma-Aldrich.
Xu S., Liu B., Yin M., Koroleva M., Mastrangelo M., Ture S., Morrell C.N., Zhang D.X., Fisher E.A, & Jin Z.G. (2016). A novel TRPV4-specific agonist inhibits monocyte adhesion and atherosclerosis. Oncotarget, 7(25), 37622-37635.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!