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Na2edta

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Na2EDTA is a chemical compound that serves as a disodium salt of ethylenediaminetetraacetic acid (EDTA). It is a common laboratory reagent used for various applications, such as metal ion chelation and buffer preparation. Na2EDTA is a white, crystalline solid with a molecular formula of Na2C10H14N2O8.

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Lab products found in correlation

Bovine serum albumin (bsa), by Avantor (5 mentions) Attune nxt flow cytometer, by Thermo Fisher Scientific (5 mentions) T bet 4b10, by BioLegend (2 mentions) Q31 378, by BD (2 mentions) Ef780, by Thermo Fisher Scientific (2 mentions) Fjk 16, by Thermo Fisher Scientific (1 mentions) L cysteine hydrochloride, by Merck Group (1 mentions) Papain, by Merck Group (1 mentions) Thermomixer, by Eppendorf (1 mentions) Ki67 sola15, by Thermo Fisher Scientific (1 mentions) Sodium deoxycholate, by Merck Group (1 mentions) Butylated hydroxytoluene, by Merck Group (1 mentions) Phospholipon 90g, by Lipoid (1 mentions) Potassium dihydrogen phosphate, by Merck Group (1 mentions) Sodium hydroxide (naoh), by Merck Group (1 mentions) Sepigel 305, by Seppic (1 mentions)

8 protocols using na2edta

1

Immunophenotyping of T Cell Subsets

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Immunofluorescence antibodies were used as purchased. The 0.1% flow staining buffer was formulated by adding 0.1% bovine serum albumin (VWR, Radnor, PA), 2mM Na2EDTA (VWR, Radnor, PA) and 0.01% NaN3 (VWR, Radnor, PA) in DI H2O. Flow cytometry was completed by following the manufacturer's guidance of the Attune NXT Flow cytometer (ThermoFisher Scientific, Waltham, MA, USA) in Arizona State University’s flow cytometry core. The following immunofluorescence antibodies were utilized in the flow cytometry studies: MHC-Tetramer: - I-A(q) bovine collagen II 271-285 GEPGIAGFKGEQGPK (NIH Tetramer Core Facility), CD4 (RM4-5, 566407, BD Biosciences), CD8 (53-6.7, 564983, BD Bioscience), CD25 (PC61, 552880, BD Biosciences), CD44 (IM7, 566200, BD Biosciences), Tbet (4B10, 644835, BioLegend), GATA3 (L50-823, 565449, BD Biosciences), RORyT (Q31-378, 564722, BD Biosciences), Foxp3 (FJK-16s, 48-5773-82, Invitrogen) and Ki67 (SolA15, 11-5698-82, Invitrogen).
Mangal J.L., Inamdar S., Suresh A.P., Jaggarapu M.M., Esrafili A., Ng N.D, & Acharya A.P. (2022). Short Term, Low Dose Alpha-ketoglutarate Based Polymeric Nanoparticles with Methotrexate Reverse Rheumatoid Arthritis Symptoms in Mice and Modulate T Helper Cell Responses. Biomaterials science, 10(23), 6688-6697.
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2

Quantification of Matrix Components

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To measure DNA, GAGs, and hydroxyproline (HYP), samples were digested, according to Kim et al. [19 (link)]. Briefly, a small piece (~ 3 mm × 3 mm) from the middle portion of each sample was taken and lyophilized for 48 hours. Three pieces of 2–3 mg of freeze-dried tissue were used as technical replicate from each meniscus and digested in 500 μL Papain digestion buffer [130 μg Papain (Sigma-Aldrich) per ml in 5 mM L-cysteine hydrochloride (Sigma-Aldrich) + 5 mM Na2EDTA (VWR)] for 24 hours at 60 °C in an Eppendorf ThermoMixer® at 300 rpm.
Spierings J., Velthuijs W., Mansoor A., Bertrand M.E., Uquillas J.A., Ito K., Janssen R.P, & Foolen J. (2022). A decellularized and sterilized human meniscus allograft for off-the-shelf meniscus replacement. Journal of Experimental Orthopaedics, 9, 116.
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3

Flow Cytometry Staining with Fixable Dye

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All the antibodies were purchased and used as is (BD biosciences, Tonbo Biosciences, BioLegend, Thermo Scientific, Invitrogen). Flow staining buffers were prepared by generating 0.1% bovine serum albumin (VWR, Radnor, PA), 2mM Na2EDTA (VWR, Radnor, PA) and 0.01% NaN3 (VWR, Radnor, PA). Live/dead staining was performed using fixable dye eF780 (ThermoFisher Scientific, Waltham, MA, USA). Flow cytometry was performed by following manufacturer’s recommendations using Attune NXT Flow cytometer (ThermoFisher Scientific, Waltham, MA, USA). The antibodies used in these studies are shown in the table below.
Mangal J.L., Inamdar S., Yang Y., Wankhede M., Shi X., Gu H., Green M., Rege K., Curtis M, & Acharya A.P. (2020). Metabolite releasing polymers control dendritic cell function by modulating their energy metabolism. Journal of materials chemistry. B, 8(24), 5195-5203.
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4

Flow Cytometry Staining Protocol

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Flow cytometry (FACS) staining buffer was prepared by generating 0.1% bovine serum albumin (VWR, Radnor, PA), 2 mM Na2EDTA (VWR, Radnor, PA) and 0.01% NaN3 (VWR, Radnor, PA). Live/dead staining was performed using fixable dye eF780 (ThermoFisher Scientific, Waltham, MA, USA). All antibodies required for staining were purchased and used as is (BD biosciences, Tonbo Biosciences, BioLegend, Thermo Scientific, Invitrogen). Flow cytometry was performed by following the manufacturer’s recommendation and guidelines set by ASU flow cytometry core using Attune NXT Flow cytometer (ThermoFisher Scientific, Waltham, MA, USA). Flowjo v10 was utilised to analyse the data obtained from the flow cytometer.
Inamdar S., Suresh A.P., Mangal J.L., Ng N.D., Sundem A., Wu C., Lintecum K., Thumsi A., Khodaei T., Halim M., Appel N., Jaggarapu M.M., Esrafili A., Yaron J.R., Curtis M, & Acharya A.P. (2023). Rescue of dendritic cells from glycolysis inhibition improves cancer immunotherapy in mice. Nature Communications, 14, 5333.
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5

Immunofluorescence Antibody Staining Protocol

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All immunofluorescence antibodies were purchased and used as is (BD biosciences, Tonbo Biosciences, Biolegend, Thermo Scientific, Invitrogen). The 0.1% flow staining buffer was prepared by mixing 0.1% bovine serum albumin (VWR, Radnor, PA), 2mM Na2EDTA (VWR, Radnor, PA) and 0.01% NaN3(VWR, Radnor, PA) in DIH2O. Fixable dye eF780 (ThermoFisher Scientific, Waltham, MA, USA) was used for live/dead staining. Flow cytometry was completed by following the manufacturer’s guidance of the Attune NXT Flow cytometer (ThermoFisher Scientific, Waltham, MA, USA) in the Arizona State University’s flow cytometry core.
Mangal J.L., Inamdar S., Le T., Shi X., Curtis M., Gu H, & Acharya A.P. (2021). Inhibition of glycolysis in the presence of antigen generates suppressive antigen-specific responses and restrains rheumatoid arthritis in mice. Biomaterials, 277, 121079.
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6

Comprehensive Immunophenotyping by Flow Cytometry

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Immunofluorescence antibodies were used as purchased. The 0.1% flow staining buffer was formulated by adding 0.1% bovine serum albumin (VWR, Radnor, PA), 2mM Na2EDTA (VWR, Radnor, PA) and 0.01% NaN3 (VWR, Radnor, PA) in DI H 2 O. Flow cytometry was completed by following the manufacturer's guidance of the Attune NXT Flow cytometer (ThermoFisher Scientific, Waltham, MA, USA) in Arizona State University's flow cytometry core. The following immunofluorescence antibodies were utilized in the flow cytometry studies: MHC-Tetramer: -I-A(q) bovine collagen II 271-285 GEPGIAGFKGEQGPK (NIH Tetramer Core Facility), CD4 (RM4-5, 566407, BD Biosciences), CD8 (53-6.7, 564983, BD Bioscience), CD25 (PC61, 552880, BD Biosciences), CD44 (IM7, 566200, BD Biosciences), Tbet (4B10, 644835, BioLegend), GATA3 (L50-823, 565449, BD Biosciences), RORyT (Q31-378, 564722, BD Biosciences), Foxp3 (FJK-16s, 48-5773-82, Invitrogen) and Ki67 (SolA15, 11-5698-82, Invitrogen).
Mangal J.L., Inamdar S., Suresh A.P., Jaggarapu M.M.C.S., Esrafili A., Ng N.D, & Acharya A.P. (2022). Short term, low dose alpha-ketoglutarate based polymeric nanoparticles with methotrexate reverse rheumatoid arthritis symptoms in mice and modulate T helper cell responses. Biomaterials science, 10(23).
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7

Leaf Catalase Activity Assay

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Fresh leaf samples were frozen in liquid nitrogen upon harvesting from both treated and control plant samples. The frozen leaves were then kept at low temperature (−80 °C) for further analyses. For each sample, 0.5 g of leaf samples was vortexed with 5 mL of extraction buffer with 50 mM K-phosphate buffer with pH of 7.6 and 0.1 mM Na2EDTA (Amresco, Dallas, TX, USA). The buffer mixture was centrifuged at 15,000 rpm for 15 min, and the supernatant fraction was used to assay various enzymes. All steps in the preparation of enzyme extracts were performed at 4 °C. Catalase (CAT) activity was determined by monitoring the declining level of H2O2, as described by Cakmak and Marschner [47 (link)].
Lu P., Magwanga R.O., Lu H., Kirungu J.N., Wei Y., Dong Q., Wang X., Cai X., Zhou Z., Wang K, & Liu F. (2018). A Novel G-Protein-Coupled Receptors Gene from Upland Cotton Enhances Salt Stress Tolerance in Transgenic Arabidopsis. Genes, 9(4), 209.
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8

Topical Drug Delivery Formulation

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rhEGF used in this study was kindly donated by Daewoong Pharmaceutical Co. Ltd. (Seoul, South Korea). Phospholipon 90G was a gift from Lipoid GmbH (Kӧln, Germany). Sodium deoxycholate and butylated hydroxytoluene were purchased from Sigma-Aldrich (St. Louis, Missouri, USA) and Sterlitamak Petrochemical Plant (Sterlitamak, Russia), respectively. Potassium dihydrogen phosphate and sodium hydroxide were obtained from Merck (Darmstadt, Germany). Sepigel 305 (Seppic, Paris, France), propylene glycol (Qingdao Aspirit Chemical Co. Ltd, Qingdao, China), Na2EDTA (Amresco, Ohio, USA), methylparaben (Clariant, Höchst,Germany), and propylparaben (Clariant, Pontypridd, UK) were of pharmaceutical grade. All other solvents and reagents were of analytical grade.
Surini S., Leonyza A, & Suh C.W. (2020). Formulation and In Vitro Penetration Study of Recombinant Human Epidermal Growth Factor-Loaded Transfersomal Emulgel. Advanced Pharmaceutical Bulletin, 10(4), 586-594.
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